MICROSCOPE SLIDE CARRYING CASSETTE

专利摘要:
microscope slide cassette, method of transport and storage device. this application concerns the imaging field and, more particularly, imaging systems, slide cassettes, and methods of processing biological samples on slides. the microscope slide carrier cassette comprises: a main body (2410) for surrounding and protecting microscope slides (2422), the main body (2410) including a first side wall and a second side wall; a plurality of partitions (2414) capable of supporting microscope slides (2422), the partitions (2414) being positioned between the first side wall (2452) and the second side wall (2454) and are vertically spaced apart from each other when a microscope slide cassette (2400) is in a vertical orientation; and a plurality of support members (2500a, 2500b) extending away from the respective partitions (2414), at least one of the support members (2500a) includes an elongated body (2508a) coupled to one of the partitions (2414) ), and a fastener (2510a) that extends upwards from the elongated body (2508a) and can limit the movement of a microscope slide (2422) positioned on the elongated body (2508a).
公开号:BR112014005007B1
申请号:R112014005007-4
申请日:2012-08-21
公开日:2020-12-15
发明作者:Raphael Hebert；Chris Todd；David Moriconi；Gregory C. Loney；Keith Moravick
申请人:Ventana Medical Systems, Inc.；
IPC主号:

专利说明:

FIELD OF THE INVENTION
[001] This application refers to the imaging field and, more particularly, imaging systems, slide cassettes, and methods for processing biological samples on slides. BACKGROUND OF THE INVENTION
[002] Identification of molecular imaging of changes in cell structures indicative of disease is often important for a better understanding in medical science. Microscopy applications are applicable to microbiology (eg, Gram stain, etc.), plant tissue culture, animal cell culture (eg, phase contrast microscopy, etc.), molecular biology, immunology (eg , ELISA, etc.), cell biology (eg, immunofluorescence, chromosome analysis, etc.), confocal microscopy, time interval and live cell imaging, serial imaging and three-dimensional imaging.
[003] In manual microscopy methods, specimen support slides are manually loaded into a microscope and viewed through microscope eyepiece lenses. In medical applications, a pathologist can inspect cellular characteristics, stained cell counts vs. untainted cells or other specimen characteristics. It often takes a lot of time to analyze a large number of specimens due to the handling time of each slide. Long processing times can delay accurate diagnosis and treatment. In addition, conventional microscopes often fail to capture high-resolution, high-quality images suitable for archiving and later use.
[004] In automated microscopy methods, digital images are often collected, viewed on high resolution monitors, shared and archived for later use. Unfortunately, conventional automatic slide processing systems often work incorrectly, including incorrect slide handling, incorrect alignment of microscope slides with optical components (eg, cameras, image capture devices, etc.), and slide breaks for microscopes. . For example, conventional robotic equipment often attaches a marker tip to a microscope slide. In this case, residual glue may be exposed on the marker. If a claw comes into contact with the edges of the blade adjacent to the marker or the edges of the marker, the exposed residual glue may stick or stick to the claw. This can result in incorrect handling of the blade. DESCRIPTION OF THE INVENTION
[005] In some embodiments, the cassette carrying a microscope slide includes a main body, a plurality of dividers and a plurality of support members. The main body is configured to surround and protect microscope slides and includes a first side wall and a second side wall. The dividers can hold microscope slides and are positioned between the first and second side walls. The dividers can be vertically spaced apart when the microscope slide cassette is in the vertical position. The support members extend away from the respective partitions and include an elongated body and a closure. The closure projects upward from the elongated body and can limit the movement of the microscope slide positioned along an elongated body.
[006] In some other embodiments, a method of transporting a microscope slide includes removing the microscope slide using a removal device without contacting an edge of a marker on the microscope slide and without contacting with a microscope slide edge adjacent to the marker. The microscope slide is transported using the removal device to a desired location. In some embodiments, the microscope slide is transported between at least two processing stations (for example, a dye unit, an automatic lamella placement device, an imaging system, an optical scanner, an optical imaging system) without entering in contact with the edge of the marker and without coming into contact with the edge of the slide.
[007] In some embodiments, a microscope slide capture device comprises a final effector that includes an elongated planar platform with an upper surface and a lower surface. A fluid line is positioned on an upper side of the platform. The top element includes a connector and a suction head. The connector is positioned on the upper side of the platform and coupled to the fluid line. The suction head is positioned on the bottom side of the platform. BRIEF DESCRIPTION OF THE DRAWINGS
[008] Non-restrictive and non-exhaustive embodiments will be described with reference to the following drawings. The same reference numbers refer to similar parts or acts throughout the various views, as long as there is no different indication.
[009] Figure 1 is a schematic illustration of an imaging system for a scanning microscope and / or other scanning device that may include various component devices used in connection with digital pathology sample scanning and imaging according to various embodiments of the system described here.
[010] Figure 2 is a schematic illustration showing an imaging device that includes a focus system in accordance with an embodiment of the system described here.
[011] Figures 3A and 3B are schematic illustrations of an embodiment of the control system that shows that the control system can include appropriate electronics.
[012] Figure 4 is a schematic illustration showing the dither focus stage in more detail according to an embodiment of the system described here.
[013] Figures 5A-5E are schematic illustrations showing an iteration of the operations of focusing operations according to the system described here.
[014] Figure 6A is a schematic illustration of a graph showing the command waveform of the dither focus optics and sharpness determinations according to an embodiment of the system described here.
[015] Figure 6B is a schematic illustration showing a graph of calculated sharpness values (Zs) for a portion of the sinusoidal wave motion of Dither standard lenses.
[016] Figures 7A and 7B are schematic illustrations showing focus determinations and adjustments of a specimen (tissue) according to an embodiment of the system described here.
[017] Figure 8 is a schematic illustration showing an example of a sharpness profile including a sharpness curve and contrast ratio for each sharpness response at multiple points that are sampled by the dither standard focusing optics according to a implementation of the system described here.
[018] Figure 9 shows a functional control loop block diagram that illustrates the use of the contrast function to produce a control signal to control the slow focus stage.
[019] Figure 10 is a schematic illustration showing the focus window being fragmented into zones in connection with the focus processing according to an embodiment of the system described here.
[020] Figure 11 shows a graphic illustration of different values of sharpness that can be obtained in points in time in an embodiment according to techniques here.
[021] Figure 12 is a flowchart showing focus processing in real time (on-the-fly) while scanning a specimen under examination according to an embodiment of the system described here.
[022] Figure 13 is a flow chart showing processing in the slow focus stage according to an embodiment of the system described here.
[023] Figure 14 is a flowchart showing image capture processing according to an embodiment of the system described here.
[024] Figure 15 is a schematic illustration showing an alternative arrangement for focus processing according to an embodiment of the system described here.
[025] Figure 16 is a schematic illustration showing an alternative focus processing arrangement according to another embodiment of the system described here.
[026] Figure 17 is a flow chart showing the processing to obtain a mosaic image of tissue on a slide according to an embodiment of the system described here.
[027] Figure 18 is a schematic illustration showing an implementation of a precision stage implementation of a precision stage (for example, a portion of stage Y) of a stage XY according to an embodiment of the system described here.
[028] Figures 19A and 19B are more detailed views of the movement stage block of the precision stage according to an embodiment of the system described here.
[029] Figure 20 shows an implementation of an entire XY composite stage according to the precision stage characteristics discussed here and includes a Stage Y, a Stage X and a base plate according to an embodiment of the system described here.
[030] Figure 21 is a schematic illustration showing a blade capture device according to an embodiment of the system described here.
[031] Figure 22A is a flow chart showing a slide capture processing according to an embodiment of the system described here in connection with a first slide.
[032] Figure 22B is a flowchart showing blade capture processing according to an embodiment of the system described here in connection with a second blade.
[033] Figures 23A and 23B show time diagrams using blade caching techniques according to the embodiments of the system described herein and illustrate time savings according to various embodiments of the system described here.
[034] Figure 24 is a schematic illustration showing a caching device according to another embodiment of the system described here.
[035] Figure 25A is a flowchart showing caching processing in connection with a first blade according to an embodiment of the system described for a composite XY two-stage caching device for blade processing.
[036] Figure 25B is a flow chart showing caching processing in connection with a second blade according to an embodiment of the system described for the XY two-stage composite caching device for blade processing.
[037] Figure 26 is a schematic illustration showing a caching device according to another embodiment of the system described here.
[038] Figure 27 is a schematic illustration showing another view of the caching device according to figure 26.
[039] Figures 28A-28J are schematic illustrations showing blade caching operations of the caching device of figures 26 and 27 according to an embodiment of the system described herein.
[040] Figure 29 is a schematic illustration showing a lighting system for lighting a blade using a light emitting diode (LED) lighting kit according to an embodiment of the system described here.
[041] Figure 30 is a schematic illustration showing a more detailed view of an embodiment for an LED lighting kit according to the system described here.
[042] Figure 31 is a schematic representation showing an exploded view of a specific implementation of an LED lighting kit according to an embodiment of the system described here.
[043] Figure 32 is a schematic illustration showing a high speed blade scanner according to an embodiment of the system described here, which can be used in connection with digital pathology imaging.
[044] Figure 33 is a schematic illustration showing a recess in a tray of the high speed blade scanner in more detail according to an embodiment of the system described here.
[045] Figure 34 is a schematic illustration showing an imaging route that starts at a first position with respect to the slide for imaging a specimen on the slide in the recess.
[046] Figures 35A and 35B are schematic illustrations showing an alternative arrangement of blades on a rotating blade holder according to another embodiment of the system described here.
[047] Figure 36 is a schematic illustration showing an imaging system in accordance with an embodiment of the system described here that includes an objective arranged to examine a specimen on a slide.
[048] Figure 37 is a flowchart showing a high-speed blade scan using a rotating tray according to an embodiment of the system described here.
[049] Figure 38 is a schematic illustration showing an optical duplication image system according to an embodiment of the system described here.
[050] Figures 39A and 39B are schematic illustrations of the optical duplication image system showing the transfer of the first tube lens and the second tube lens in front of the image sensor according to an embodiment of the system described here.
[051] Figure 40 is an isomeric left, front and top view of an imaging system, according to one embodiment.
[052] Figure 41 is an isometric left, front and top view of an imaging system in figure 40. An external protective box is shown removed.
[053] Figure 42 is a right, rear, isometric side view of the top of the imaging system of figure 40 with the shown housing removed.
[054] Figure 43 is a top plan view of the imaging system in Figure 40.
[055] Figure 44 is an isomeric view of a gripping device, according to an embodiment.
[056] Figure 45 is a top plan view of the gripping device of Figure 44.
[057] Figure 46 is a cross-sectional view of the gripping device taken along line 46-46 in figure 45.
[058] Figure 47 is an isomeric view of the gripping device in Figure 44.
[059] Figure 48 is a detailed view of the gripping device in Figure 46.
[060] Figure 49 is a right lateral isomeric view from the front, from the top of the blade holder device.
[061] Figure 50 is a left, top, rear isomeric side view of the blade holder device of figure 49.
[062] Figure 51 is a front elevation view of the blade holder in Figure 49.
[063] Figure 52 is a cross-sectional view of the blade holder taken along a line 52-52 of figure 51. A grip device is positioned to insert a blade into the blade holder.
[064] Figure 53 is a detailed view of the blade holder in Figure 52.
[065] Figure 54 is a detailed view of one end of an upper partition.
[066] Figure 55 is a cross-sectional view of the blade holder taken along a line 53-53 in figure 51.
[067] Figure 56A is a front elevation view of a blade positioned above an upper partition of a blade holder.
[068] Figure 56B is a front elevation view of the blade that rests on the upper part of Figure 56A. DESCRIPTION OF ACCOMPLISHMENTS OF THE INVENTION
[069] Figure 1 is a schematic illustration of an imaging system 5 of a scanning microscope and / or other scanning device that may include various component devices used in connection with digital pathology sample scanning and imaging according to various embodiments of the system described here. The imaging system 5 may include an imaging device with a focusing system 10, a blade stage 20 system, a blade caching system 30, and a lighting system 40, among other component systems 50, as per the followed in more detail here. It was also found that the System described here can be used in connection with microscope slide scanning instrument architectures and techniques for image capture, image splicing and magnification as described in the US Patent App. Pub. No. 2008 / 0240613 A1 by Dietz et al., Entitled “Digital Microscope Slide Scanning System and Methods,” which is incorporated by reference, including features in connection with reconstitution of an image with a magnification without substantial loss of accuracy and display or storage of the image reconstituted.
[070] Figure 2 is a schematic illustration showing an imaging device 100 of an optical scanning microscope and / or other appropriate imaging system that includes components of a focusing system for obtaining focused images of a tissue sample 101 and / or another object disposed on a blade according to an embodiment of the system described herein. The focusing system described here provides the determination of the best focus for each snapshot when a snapshot is captured, which can be called “on-the-fly focus.” The devices and technique provided here enable significant reductions in the time required to form a digital image of an area on a pathology slide. The System described here integrates steps from the two-step method of conventional systems and essentially eliminates the time required for pre-focusing. The System described here provides the creation of a digital image of a specimen on a microscope slide using real-time processing to capture snapshots in which the total time to capture all snapshots is less than the time required by the method that uses a pre-determination of focus points for each snapshot before capturing the snapshots.
[071] Imaging device 100 may include an imaging sensor 110, such as a charge-coupled device (CCD) and / or complementary metal oxide semiconductor (CMOS) sensor that can be part of a camera 111 that captures images of digital pathology. Imaging sensor 110 can receive light transmitted from a microscope objective 120 transmitted through a tube lens 112, a beam separator 114 and which includes other components of a transmitted light microscope such as a condenser 116 and a light source 118 and / or other appropriate optical components 119. Microscope objective 120 can be corrected to infinity. In one embodiment, beam splitter 114 can provide approximately 70% attribution of the light beam source directed to image sensor 110 and the remaining portion of approximately 30% directed along a route to the dither pattern focusing stage 150 and focus sensor 160. The tissue sample 101 being imaged can be arranged in a movement stage with XY 130 movement that can be moved in the X and Y directions and can be controlled as discussed here. Focus stage 140 can control the movement of microscope objective 120 in the Z direction to focus an image of tissue 101 that is captured by image sensor 110. Focus stage 140 may include a motor and / or other device suitable for moving the microscope objective 120. A dither 150 focusing stage and a 160 focus sensor are used to provide fine focus control for real-time focusing according to the system described here. In various embodiments, the focus sensor 160 can be a CCD sensor and / or CMOS sensor.
[072] The focus stage with dither standard 150 and the focus sensor 160 provide real-time focusing according to the sharpness values and / or other measurements that are quickly calculated during the imaging process in order to obtain a better focus for each snapshot of image when it is captured. As will be discussed in detail here, the focusing stage with dither pattern 150 can be moved at a frequency, for example, in a sinusoidal movement that is independent of and exceeds the practicable movement frequency for the slower movement of the microscope objective 120. Multiple measurements are made by the focus sensor 160 of focus information for tissue views in the range of motion of the focus stage with dither pattern 150. The focus electronics and control system 170 may include focus sensor and stage control electronics of focus with dithering 150, a master clock, electronic control of stage of slow focus 140 (Direction Z), stage with movement XY 130, and other components of a realization of a system according to the techniques discussed here. The focus electronics and control system 170 can be used to perform sharpness calculations using information from the focusing stage with dither standard 150 and focus sensor 160. Sharpness values can be calculated using at least a portion of a curve sinusoidal motion-defined with dither pattern. The focus electronics and control system 170 can use information to determine the position for the best focus image of the tissue and command the slow focus stage 140 to move the microscope objective 120 to the desired position (along the Z axis as shown ) to obtain the best focus image during the imaging process. The control system 170 can also use the information to control the speed of the stage with XY 130 movement, for example, the movement speed of stage 130 in the Y direction. In one embodiment, sharpness values can be computed by differentiating values of neighboring pixels, scanning them and adding those values to form a score. Various algorithms for determining sharpness values will also be covered here.
[073] In various embodiments according to the system described here, and according to the components discussed here, a device for creating a digital image of a specimen on a microscope slide includes: a microscope objective that is corrected To infinity; a beam splitter; a camera focusing lens; a high resolution camera; a sensor focus lens group; a focus stage with a dither pattern; a focus sensor; an approximate (slow) stage of focusing; and focus electronics. The device can allow the lens to focus and capture each snapshot through the camera without the need to pre-determine a focus point for all snapshots before the snapshots are captured, and the total time to capture all snapshots. snapshots is less than the time required through a system request for a pre-determination of focus points for each snapshot before the snapshots are captured. The System can include computer controls for: i) determining a first focus point on the tissue to establish a nominal focus plane by moving the approximate focus stage across the entire z range and monitoring sharpness values; ii) positioning the tissue in x and y to start in a corner of an area of interest; iii) establishment of the fine focus stage with dither pattern to move, the focus stage with dither pattern being synchronized to a master clock that also controls the speed of XStage Y; iv) command of the stage to move from the frame to an adjacent frame, and / or v) production of a trigger signal to obtain a frame in the image sensor and activate a light source to create a pulse of light.
[074] In addition, according to another embodiment, the system described here can provide a computer-implemented method for creating a digital image of a specimen on a microscope slide. The method may include determining a scanning area comprising a region of the microscope slide that includes at least a portion of the specimen. The scan area can be divided into a plurality of snapshots. Snapshots can be captured using a microscope lens and camera, in which focusing the lens and microscope and capturing each snapshot through the camera can be conducted for each snapshot without the need to pre-determine a focal point for all snapshots before snapshots are captured. The total time to capture all snapshots can be less than the time required by a method that prompts a pre-determination of the spot point for each snapshot before capturing the snapshots.
[075] Figure 3A is a schematic illustration of an embodiment of the focus electronics and control system 170 including focus electronics 161, a master clock 163 and stage control electronics 165. Figure 3B is a schematic illustration of an embodiment of focus electronics 161. In the illustrated embodiment, focus electronics 161 may include appropriate electronics such as a suitably fast A / D converter 171 and a programmable field gate arrangement (FPGA) 172 with a microprocessor 173 that can be used to perform sharpness calculations. The A / D converter 171 can receive information from the focus sensor 160 which is coupled to the FPGA 172 and microprocessor 173 and is used to emit sharpness information. The master clock included in system 170 can supply the master clock signal to focus electronics 161, stage control electronics 165, and other system components. Stage control electronics 165 can generate control signals used to control slow focus stage 140, XY 130 moving stage, dither 150 focusing stage, and / or other control and information signals, as follows covered here. The FPGA 172 can provide a clock signal to the focus sensor 160, among other information. Laboratory measurements show a sharpness calculation in a 640 x 32 pixel frame can be made in 18 microseconds, fast enough for the proper operation of the system described here. In one embodiment, the focus sensor 160 may include a monochrome CCD camera in 640 x 32 strip mode, as discussed hereinafter.
[076] The scanning microscope can achieve a 1D or 2D set of pixels including contrast information and / or intensity information in RGB or some other colored space as discussed below. The system finds the best focus points over a large field, for example on a 25 mm x 50 mm glass slide. Many commercial systems sample the scene produced by a 20x, 0.75 NA lens for a microscope with a CCD series. Given the NA of the objective and capacitor of 0.75 and wavelength of 500 nm the lateral resolution of the optical system is approximately 0.5 microns. To sample this resolution element at the Nyquist frequency, the pixel size in the object is approximately 0.25 microns. For a 4 Mpixel camera (for example, a False Falcon 4M30 / 60), which operates at 30 fps, with a pixel size of 7.4 microns the magnification from the object to the imaging camera is 7.4 / 0.25 = 30x. Therefore, a 2352 x 1728 frame can cover an area of 0.588mm x 0.432mm on the object, which equates to approximately 910 frames for a typical fabric section defined as 15 mm x 15 mm in area. The system described here is used where the spatial variation of tissue in the focus dimension is much less than the frame size on the object. Variations in focus, in practice, occur over greater distances and much of the focus adjustment is done to correct slopes. These slopes are generally in the range of 0.5 - 1 micron per frame size on the object.
[077] The time to result from current scanning systems (for example, a BioImagene iScan Coreo system) is approximately 3.5 minutes for pre-scanning and scanning a 20x 15 mm x 15 mm field and approximately 15 minutes for a 40x field scan of 15 mm x 15 mm. The 15 mm x 15 mm field is scanned by rotating 35 frames in 26 steps. Scans can be done unidirectionally with a reconstitution time of 1 second. The time to scan using a technique according to the system described here can be approximately 5 seconds to find the nominal focus plane, 1.17 seconds per step (25 steps), for a total of 5 + 25 x (1.17 + 1) = 59.25 seconds (approximately 1 minute). This is a considerable saving of time compared to conventional methods. Other embodiments of the systems described here may allow even faster focusing times, but there may be a limitation in the amount of light required for short illumination times in order to avoid motion blur in continuous scanning. Pulsation or intermittent illumination of the light source 118, which can be an LED light source as discussed below, to enable high peak lighting can minimize this issue. In one embodiment, the pulsation of the light source 118 can be controlled by the focus electronics and control system 170. In addition, the operation of the bi-directional system eliminates the saving of reconstitution time in approximately 25 seconds for a 20x scan resulting in a scan time of 35 seconds.
[078] Note that the components used in connection with the focus electronics and control system 170 can also more generally be called electrical components used to perform a variety of different functions in connection with embodiments of the techniques described here.
[079] Figure 4 is a schematic illustration showing the focus stage with dither pattern 150 in more detail according to an embodiment of the system described here. The dither pattern focus stage 150 can include a dither pattern focus lens 151 that can be moved by one or more actuators 152a, b, such as voice coil actuators, and that can be mounted on a rigid housing 153. In one embodiment, the lens can be an achromatic lens with a focal length of 50 mm as commercially available, see for example Edmund Scientific, NT32-323. Alternatively, the focusing lens with dither pattern 151 can be constructed of plastic, aspherical material and in a shape like that in which the lens weight is reduced (extremely low mass). A flexural structure 154 can be attached to the rigid housing 153 and attached to a rigid grounding point and can only allow translational movement of the focusing lens with dither pattern 151, for example, short distances of approximately 600-1000 microns. In one embodiment, the flexural structure 154 can be constructed with appropriate stainless steel sheets approximately 0.010 '' thick in the flexing direction and form a connection of four bars. The flexural texture 154 can be designed from a suitable spring steel in a work effort far from its fatigue limit (factor 5) to operate for many cycles.
[080] The mass of motion of the focusing lens with dither pattern 151 and flexural structure 154 can be designed to provide approximately a first mechanical resonance of 60 Hz or more. The movement mass can be monitored with a suitable high bandwidth position sensor (for example,> 1 kHz) 155, such as a capacitive sensor or eddy current sensor to provide feedback to the control system 170 (see Figure 2 ). For example, KLA Tencor's ADE division manufactures a 5 mm 2805 capacitive sensor probe with a 1 kHz bandwidth, 1 mm measurement range and 77 nanometer resolution suitable for this application. The focus with dither pattern and control system, as represented by functionality included in the 170 system, can maintain the amplitude of the focus lens with dither pattern 151 for a prescribed focus range. The focus with dither pattern and control system may depend on well-known controlled gain oscillator circuits. When operated in resonance, the focus lens with dither pattern 151 can be activated at low current dissipating low power in the voice coil windings. For example, with the use of a BEI Kimco LAO8-10 actuator (winding A) the average currents can be less than 180 mA and the dissipated power can be less than 0.1 W.
[081] Note that other types of movement of the Dither standard lenses and other types of actuators 152a, b can be used in connection with various embodiments of the system described here. For example, piezoelectric actuators can be used as actuators 152a, b. In addition, the movement of Dither standard lenses can be movement at other resonant frequencies that remains independent of the movement of the microscope objective 120.
[082] Sensor 155, like the capacitive sensor mentioned above, can be included in an embodiment according to the techniques described here, can provide feedback as to where the focusing lens with dither pattern is positioned (for example, with respect to where sinusoidal or cycle corresponding to the movements of the lens). As per cera___2 described here, a determination can be made as to which picture frame picture frame obtained using the focus sensor produces the best sharpness value. For this table, the position of the focusing lens with dither pattern can be determined with respect to the sine wave position as indicated by sensor 155. The position as indicated by sensor 155 can be used by 170 control electronics to determine an appropriate setting for the focus stage 140. For example, in one embodiment, the movement of the microscope objective 120 can be controlled by a slow step motor from the slow focus stage 140. The position indicated by sensor 155 can be used to determine an amount corresponding motion (and corresponding control signal (s) to position the microscope objective 120 in the best focus position in the Z direction. The control signal (s) can be transmitted to the slow focus stage step 140 to enable any necessary repositioning of microscope objective 120 in the best focus position.
[083] Figures 5A-5E are schematic illustrations showing an iteration of the focusing operations according to the system described here. The figures show the image sensor 110, the focus sensor 160, the focus stage with dither standard 150 with a Dither standard lens and the microscope objective 120. Fabric 101 is illustrated moving on the Y axis, ie, on the Stage with XY 130 movement, while focus operations are performed. In one example, the dither 150 focusing stage can move the Dither standard lenses at a desired frequency, such as 60 Hz or more (for example, 80 Hz, 100 Hz), although in other embodiments, the system described here can also operate with the standard Dither lens that moves at a lower frequency (for example, 50Hz) according to the applicable circumstances. The XY 130 moving stage can be commanded to move, for example, in the Y direction, from the frame to the adjacent frame. For example, stage 130 can be commanded to move at a constant 13 mm / second which for a 20x objective corresponds to an acquisition rate of approximately 30 frames / second. As the focus stage with dither 150 pattern and stage with XY 130 movement can be locked in phase, the focus stage with dither 150 pattern and sensor 160 can perform 60 focus calculations per second, or operate with 120 bi-focus points. directional (reading in the up and down movement of the sine wave) per second or 4 focus points per frame. For a frame height of 1728 pixels, this is equivalent to a focus point for every 432 pixels or for the 20x lens for every 108 microns. Since the XY 130 moving stage is in motion, the focus point must be captured in a very short time, for example 330 μsec (or less), to maintain the minimum variation in the scene.
[084] In various embodiments, as discussed below, these data can be stored and used to extrapolate the next frame focus position, or alternatively, extrapolation can be used and the last focus point is used for the position focus of the active frame. With a dithered frequency of 60 Hz and a frame rate of 30 frames per second, the focus point is taken up to 1/4 of a frame from the center of the frame with the captured image. Fabric heights generally do not change enough in 1/4 of a frame to make this focus point inaccurate.
[085] A first focus point can be found on the fabric to establish the nominal or reference plane 101 '. For example, the reference plane 101 'can be determined by the initial movement of the microscope objective 120, using the slow focus stage 140, across the entire Z range I say + 1 / -1 mm and monitoring sharpness values. Once the reference plane 101 'is found, the fabric 101 can be positioned at X and Y to start at a corner and / or other specific location in the area of interest, and the dither 150 focusing stage is adjusted to move and / or otherwise move the focus stage with dither pattern 150 will continue to be monitored, starting in figure 5A.
[086] The focus stage with dither pattern 150 can be synchronized to a master clock in the control system 170 (see Figure 2), which can also be used in connection with the speed control of the XY 130 moving stage. For example , if the focus stages with dither 150 pattern shifted through a sinusoidal movement of 0.6 millimeter pv (peak to valley) at 60 Hertz, assuming a 32% duty cycle to use the more linear sinusoidal range, 8 points should be collected across the focus range over a 2.7 msec period. In figures 5B-5D, the dither pattern focusing stage 150 moves the Dither pattern lenses in a sinusoidal motion and focus samples are taken along at least a portion of the sinusoidal curve. Focus samples should therefore be taken every 330 μsec or at a rate of 3 kHz. With a magnification of 5.5x between the object and the focus sensor 160, a movement in the lens with a Dither standard of 0.6 mm p-v is equivalent to a movement of 20 micron p-v in the objective lens. This information is used to transmit the position at which maximum sharpness is computed, ie, the best focus, for the slowest stepper motor in the slow focus stage 140. As shown in figure 5E, the slow focus stage 140 is controlled to move the microscope objective 120 to the best focus position (illustrated by the 120 'motion range) in time for the image sensor 110 to capture the best focus image 110' of the tissue 101 area of interest. In one embodiment, the image sensor 110 can be triggered, for example, by the control system 170, to capture an image after a specific number of cycles of the lens movement with the Dither pattern. The XY 130 movement stage moves to the next frame, the cyclic movement of the lens with the Dither pattern in the focus stage with the dither pattern 150 continues, and the focusing operations of figures 5A-5E are repeated. Sharpness values can be calculated at a rate that does not hinder the process, for example, 3 kHz.
[087] Figure 6A is a schematic illustration of a graph 200 showing the command waveform of the focus optics with dither pattern and sharpness determinations according to an embodiment of the system described here. In an embodiment based on the times covered in connection with the example in Figures 5A-5E: T = 16.67 msec, / * lens sinusoid period with Dither pattern if the lens resonates at 60 Hz * / F = 300 μm, / * range positive of focus values * / N = 8, / * number of focus points obtained in the period E * / Δt = 330 μsec, / * focus point samples obtained every 330 μsec * / E = 2.67 msec, / * the period during which the N focus points are obtained * / Δf = 1.06 μm at the center of the focus path. / * step size of the focus curve * /
[088] So with this 32% duty cycle, 8.48 μm (8 x 1.06 μm = 8.48 μm) is sampled through focus processing.
[089] Figure 6B is a schematic illustration showing a graph 210 of calculated sharpness values (Zs) for a portion of the lens sine wave motion with the Dither pattern shown in graph 210. The position (z) for each plane of focus sampled as a function of each point i is given by EQUATION 1:

[090] Window reduction of a CCD camera can provide a high frame rate suitable for the system described here. For example, Dalsa of Waterloo, Ontario, Canada makes the Genie M640-l / 3 640 x480 monochrome camera. The Genie M640-l / 3 operates at 3,000 frames / second at a frame size of 640 x 32. The pixel size in the CCD set is 7.4 microns. At 5.5x magnification between the object and the focus plane, a focus pixel is equivalent to approximately 1.3 microns in the object. Although some calculation of the average of approximately 16 object pixels (4x4) per pixel of focus may occur, sufficient contrast shift from high spatial frequency is preserved to obtain good focus information. In one embodiment, the position of best focus can be determined according to the peak value of the graph of sharpness calculations 210. In additional embodiments it turns out that other focus calculations and techniques can be used to determine the position of best focus according to other measures including the use of a contrast measure, as discussed below here.
[091] Figures 7A and 7B are schematic illustrations showing determinations of focus and adjustments of a specimen (tissue) according to an embodiment of the system described here. In figure 7A, illustration 250 is a view of the specimen shown in approximate image frames in connection with movement of the specimen along the Y Axis according to the movement of the XY 130 moving stage discussed here. A crosspiece or passage through the specimen in connection with movement of the specimen along the Y axis (for example, according to X Stage Y movement) is illustrated in 250. Illustration 250 'is an enlarged version of a portion of the illustration 250. A frame in illustration 250 'is called dtp, with reference to a definitive tissue point in the specimen. In the example in illustration 250 ', a specimen perimeter is shown during the scan, multiple focus calculations are performed according to the system described here. Table 251 illustrates the fact that a best focus determination is made after 4 focus calculations (shown as focus positions 1, 2, 3 and 0 *) have been performed in connection with specimen, although further focus calculations can be performed in connection with the system described here. Figure 7B shows a schematic illustration 260 showing a graph of the position of the Z axis of the microscope objective in relation to the position of the Y Axis of the specimen being examined. The illustrated position 261 shows the position determined along the Z axis for adjusting the microscope objective 120 in order to obtain the best focus according to an embodiment of the system described herein.
[092] Note that the system described here provides important advantages over conventional systems such as those described in US Patent No. 7,576,307 and 7,518,642, which are incorporated by reference in which the entire microscope objective is moved through the focus in a sinusoidal or triangular pattern. The system provided here is advantageous in that it is suitable for use with a microscope objective and a monitoring stage that are heavy (especially if other objectives were added via a turret) and cannot be moved at the higher frequencies described using optics with dither pattern. The Dither standard lens described here may have an adjusted mass (for example, be designed lighter, less glass) and the imaging demands on the focus sensor are less than those imposed by the microscope objective. Focus data can be taken at high rates as described here, to minimize scene variation when computing sharpness. By minimizing scene variation, the system described here reduces discontinuities in the sharpness metric when the system moves in and out of focus while the tissue is moving under the microscope objective. In conventional systems, such discontinuities add noise to the calculation of better focus.
[093] Figure 8 is a schematic illustration 300 showing an example of a sharpness profile, produced from movement through focus positions, including a sharpness curve and contrast ratio for each sharpness response at multiple points that are sampled using the dither standard focusing optics according to an embodiment of the system described here. Graph 310 shows lens width with Dither pattern in micrometers on the X axis and sharpness units along the Y axis. As illustrated, the lens movement with Dither pattern can be centered on representative points A, B, C, D and E; however, it was found that the computations described that can be applied to each of the points on the sharpness curve. The sharpness response produced from the focus sensor 160, for a half-cycle of the sinusoidal lens with the Dither pattern, when the movement of the lens with the Dither pattern is centered at each of the points A, B, C, D and E , is shown respectively in graphs 310a-e. Based on this, the contrast ratio for each of the sharpness responses that have one of the corresponding A-E points is computed according to: Contrast function = (max - min) / (max + min). In connection with the contrast function determined for one of the AE points (for example, on which Dither pattern lens movement is centered) and one of the corresponding sharpness response curves 310a-e, max represents the maximum sharpness value obtained from sharpness response curve and min represents the minimum sharpness value obtained from the sharpness response curve. The resulting contrast function graph 320 is shown below the sharpness curve graph 310 and graphs contrast ratio values corresponding to the lens movement with Dither pattern according to the lens amplitude with Dither pattern. The minimum level of the contrast function in Graph 320 is a better focus position. Based on the contrast function and determining the best focus position, a control signal can be generated which is used to control the slow focus stage 140 to move the microscope objective 120 to a better focus position before the image 110 capture image 110 '.
[094] Figure 9 shows a block diagram of functional control loop 350 that illustrates the use of the contrast function to produce a control signal to control the slow focus stage 140. Ud can be considered as a disturbance to the loop focus control and can represent a blade slope or change in fabric surface heights, for example. Functional block 352 shows the generation of sharpness vector information that can be generated by the focus sensor 160 and communicated to the focus electronics and control system 170. Functional block 354 shows the generation of a contrast number (for example, contrast function) at the point at which the Dither standard lens is sampling the focus. This contrast number is compared to a specific point or reference value (Ref) produced in an initial stage where the best focus had previously been established. The error signal produced from this comparison with appropriate applied gain K1 (in function block 356) corrects the slow focus motor that acts (in function block 358) to keep the scene in focus. It has been found that one embodiment can adjust the position of the microscope objective 120 according to a minimum value or limit amount of movement. In this way, such an embodiment can avoid making adjustments less than the limit value.
[095] Figure 10 is a schematic illustration showing the focus window 402 being fragmented into zones in connection with focus processing according to an embodiment of the system described here. In the illustrated embodiment, the focus window is subdivided into 8 zones (402 '); however, less than or more than 8 zones can be used in connection with the system described here. A first subset of the zones can be within a snapshot n and a second subset of zones can be within a snapshot n + 1. For example, Zones 2, 3, 4, 5 are within the 404 image frame with the image captured in time t1. Zones 6 and 7 can be completely within the next image frame to be photographed when the XY 130 moving stage crosses from the bottom to the top in the figure and / or Zones 0 and 1 can be completely within the next image frame to be photographed when stage 130 crosses from the top to the bottom of the figure. Focus positions 0, 1, 2, and 3 can be used to extrapolate a better focus position to the next frame photographed at position 0 *. Fabric coverage can be established, for example, by running a serpentine pattern that covers the entire area of interest.
[096] The rectangular window 404 of the image sensor can be oriented in the direction of stage 130, just as a column of frames acquired during imaging is aligned with the rectangular focus window 402. The size of the object in the 406 image frame, using, for example, a Dalsa 4M30 / 60 CCD camera, is 0.588 mm x. 0.432 mm using a 30x tube lens magnification. The set size can be (2352 x 7.4 micron / 30) x (1720 x. 7.4 micron / 30). The widest dimension 406 of image frame (0.588 mm) can be oriented perpendicularly to the focus window 402 and allows the minimum number of columns to cross over a section of fabric. The focus sensor is 0.05 mm x. 0.94 mm using a 5x magnification on the focus leg. The rectangular focusing window 402 can be (32 x. 7.4 micron / 5.0) x (640 x 7.4 micron / 5.0). Therefore, frame 402 of the focus sensor can be approximately 2.2x higher than frame 404 of the image sensor, and can be advantageously used in connection with a look-ahead focusing technique involving multiple zones, as discussed below. According to an embodiment of the system described here, determination of best focus 120 can be performed per second, with a sharpness calculation done every 333 μsec, resulting in 8 sharpnesses calculated over 2.67 msec equal to a duty cycle of approximately 32% for a dither 8.3 msec part-time lens movement with Dither standard.
[097] A sharpening metric for each zone can be computed and stored. When computing a sharpening metric for a single focus point using multiple zones, the sharpening metric can be determined for each zone and combined, for example, such as by adding all the sharpening metrics for all zones considered in such a single point. An example of sharpness computation is shown in EQUATION 2 (for example, based on using a windowed camera for a 640 x 32 strip). For row i, dimension n up to 32, and column j, dimension m up to 640 / z, where z is the number of zones, the sharpness for each zone can be represented by EQUATION 2:
where k is an integer between or equal to 1 and 5. Other sharpening metrics and algorithms can also be used in connection with the system described here. When the XY 130 moving stage is moving along the Y axis, the system acquires sharpness information for all zones 0-7 in focus window 402. It is desirable when stage 130 is moving to know how the section heights of fabric are varying. When computing a sharpness curve (maximum sharpness being the best focus), by varying the focus height, Zones 6 and 7, for example, can provide information before the movement of the next frame where the next plane with the best focus is positioned. If major changes in focus are anticipated by early reading, stage 130 may be delayed to provide less spaced points to better track the height transition.
[098] During the scanning process, it can be advantageous to determine whether the system is moving from a white space (without fabric) to a darker space (fabric). Through the computation of sharpness in zones 6 and 7, for example, it is possible to predict whether this transition is about to happen. During the sweep of the column, if zones 6 and 7 show increased sharpness, the XY 130 moving stage can be commanded to slow down to create less spaced focus points on the fabric perimeter. If, on the one hand, movement from high sharpness to low sharpness is detected, then cera___2 it is possible to determine that the scanner's view is entering a white space and it may be desired to slow down stage 130 to create less spaced focus points on the fabric perimeter. In areas where these transitions do not occur, stage 130 can be commanded to move at higher constant speeds in order to increase the overall blade sweep performance. This method can advantageously allow a quick scan of tissue. According to the system described here, snapshots can be taken while focusing data is collected. In addition, all focus data can be collected in a first scan and stored and snapshots can be taken at points of best focus during a subsequent scan. One embodiment may use contrast ratio or function values in a similar manner to that as described herein with sharpness values to detect changes in focus and correspondingly determine transitions into, or out of, areas containing tissue or white space.
[099] For example, for a 15 mm x 15 mm 20x scan, in the frame size of 0.588 x 0.432 mm, there are 26 columns of data, each column has 35 frames. At an imaging rate of 30 fps each column is traversed in 1.2 seconds or a scan time of approximately 30 seconds. As the focus sensor 160 computes 120 (or more) focus points 120 per second, the system described here can obtain 4 foci per frame (120 foci / second divided by 30 fps). At an imaging rate of 60 fps, the scan time is 15 seconds and 2 foci per frame (120 focus / second divided by 60 fps).
[0100] In another embodiment, a color camera can be used as a focus sensor 160 and a metric chroma can be determined alternatively and / or in addition to the sharpness contrast metric. For example, a Dalsa color version of the 640 x 480 Genie camera can be properly used as a focus sensor according to this embodiment. Metric chroma can be described as color intensity with respect to the brightness of a white lit in a similar way. In the form of an equation (EQUATIONS 3A and 3B), chroma (C) can be a linear combination of color measures R, G, B: CB = -37,797 xR - 74,203xG + 112 x B (EQUATION 3A) CR = 112 x R - 93,786 x G - 18,214 x B (EQUATION 3B)
[0101] Note for R = G = B, CB = CR = 0. A value for C, which represents total chroma, can be determined based on CB and CR (for example, such as by adding CB and CR).
[0102] When the XY 130 Moving Stage is moving along the Y axis, the focus sensor 160 can acquire color information (R, G, B), as in a bright field microscope. It is desired when the stage is moving, to know how the tissue section heights are varying. The use of RGB color information can be used as with the contrast technique, to determine if the system is moving from a white space (without fabric) to a colored space (fabric). Through chroma computation in zones 6 and 7, for example, it is possible to predict whether this transition is about to happen. If, for example, a very small chroma is detected, then C = 0 and it will be possible to recognize that no tissue perimeter is being addressed. However, during scan of the focus column, if Zones 6 and 7 show increased chroma then stage 130 can be commanded to slow down to create less spaced focus points on the fabric perimeter. If, on the other hand, a movement from high chroma to low chroma is detected, then it will be determined that the scanner is entering a white space, and it may be desired to slow down stage 130 to create less spaced focus points on the fabric perimeter. In areas, where these transitions do not occur, stage 130 can be commanded to move at constant higher speeds to increase the overall blade sweep performance.
[0103] In connection with the use of sharpness values, contrast ratio values and / or chroma values to determine when the field of view or frame approach (s) is entering or leaving a slide area with tissue, processing variations may be made. For example, when an area with tissue enters from white space (for example, between two areas of tissue), the movement in the Y direction may be reduced and a number of focus points obtained may also increase. When viewing white space or an area between tissue samples, movement in the Y direction may be increased and fewer focus points determined until movement over an area containing tissue is detected (for example, such as increased chroma and / or sharpness values) ).
[0104] Figure 11 shows a graphic illustration of different values of sharpness that can be obtained in points in time in an embodiment according to techniques described here. The top portion 462 includes a curve 452 corresponding to a sine wave half-cycle (e.g., half a single cycle or peak period) of the dither pattern lens movement. The X axis corresponds to lens amplitude values with a Dither pattern during this cycle and the Y axis corresponds to sharpness values. Each point, such as point 462a, represents a point at which a frame is obtained using the focus sensor where each frame is obtained in a lens amplitude with a Dither pattern represented by the point's X-axis value and has a value sharpness represented by the point's Y Axis value. Element 465 in the lower portion 464 represents a fitted curve for each set of sharpness values obtained as shown in the upper portion 462 for the illustrated data points.
[0105] Figure 12 is a flowchart 500 that shows real-time focus processing while scanning a specimen under inspection according to an embodiment of the system described here. In a step 502, a nominal focus plane or reference plane can be determined for the specimen being examined. After step 502, processing continues until one to step 504 where a lens with a Dither standard, according to the system described here, is set to move at a specific resonant frequency. After step 504, processing continues to step 506 where the Stage with XY movement is commanded to move at a specific speed. Note that the order of steps 504 and 506, as with other processing steps referred to here, can be appropriately modified according to the system described here. After step 506, processing takes place up to step 508 where sharpness calculations for focus points with respect to the specimen being examined are carried out in connection with the movement (eg sinusoidal) of the lens with Dither standard according to with the system described here. Sharpness calculations may include the use of contrast, chroma and / or other appropriate measures as discussed below.
[0106] After step 508, processing takes place until step 510 where a position of best focus is determined for the position of a microscope objective used in connection with an image sensor to capture an image according to the system described here. After step 510, processing takes place until step 512 where a control signal concerning the best focus position is sent to a slow focus stage that controls the position (x to Z) of the microscope objective. Step 512 may also include sending a trigger signal to the camera (for example, image sensor) to capture an image of the specimen portion under the lens. The trigger signal can be a control signal that makes it possible to capture the image by the image sensor such as, for example, after a specific number of cycles (for example, as related to lens movement with dither pattern). After step 512, processing takes place until a test step 514 where it is determined whether the speed of the Stage with XY movement, which retains the specimen under scan, should be adjusted. The determination can be made according to advance reading processing techniques using sharpness and / or other information from multiple zones in a field of focus of view as discussed in detail here. If, in test step 514, it is determined that the speed of XStage Y should be adjusted, then processing takes place until a step 516 where the speed of the Stage with XY movement will be adjusted. After step 516, processing takes place back to step 508. If, in test step 514, it is determined that no adjustment to the speed of the Stage with XY movement should be made, then processing takes place until a test step 518 where it is determined whether focus processing should continue. If processing has to continue then processing will return to step 508. Otherwise, if processing does not continue (for example, scanning of the current specimen is complete), then focus processing will be completed and processing will be complete.
[0107] Figure 13 is a flow chart 530 showing processing in the slow focus stage according to an embodiment of the system described here. In a 532 step, the slow focus stage, which controls a position (for example, along the Z axis) of a microscope objective, receives a control signal with information for adjusting a position of the microscope objective you are examining a body of evidence. After step 532, processing takes place until step 534 where the slow focus stage adjusts the position of the microscope objective according to the system described here. After step 534, processing takes place until a wait step 536 where the slow focus stage expects to receive another control signal. After step 536, processing takes place back to step 532.
[0108] Figure 14 is a flow chart 550 showing image capture processing according to an embodiment of the system described here. In a 552 step, a camera's image sensor receives a trigger signal and / or other instruction that triggers processing to capture an image of a specimen under microscope examination. In various embodiments, the trigger signal can be received from a control system that controls the activation of the image sensor's image capture processing after a specific number of motion cycles of a Dither standard lens used in image processing. focus according to the system described here. Alternatively, the trigger signal can be provided based on a sensor position in the Stage with XY movement. In one embodiment, a position sensor may be a Renishaw Linear Encoder model No. T1000-10A. After step 552, processing takes place up to step 554, where the image sensor captures an image. As discussed in detail here, the image captured by the image sensor may be in focus in connection with the operation of a focusing system according to the system described here. Captured images can be stitched according to other techniques referenced here. After step 554, processing takes place until step 556 where the image sensor expects to receive another trigger signal. After step 556, processing takes place back to step 552.
[0109] Figure 15 is a schematic illustration 600 showing an alternative arrangement for processing the focus according to an embodiment of the system described here. A window focus sensor may have a frame of view (FOV) 602 field that can be tilted or otherwise positioned to sweep diagonally a range substantially equal to the width of the FOV 604 imaging sensor frame. As described here, the window may be tilted in the direction of travel. For example, the FOV 602 frame of the inclined focus sensor can be rotated 45 degrees which should have an effective width of 0.94 x 0.707 = 0.66 mm in the object (fabric). The FOV 604 frame of the imaging sensor can have an effective width of 0.588 mm, so when the Stage with XY movement that retains tissue moves under the lens, the inclined FOV 602 frame focus sensor sees the corners of the range observed by the sensor of image. In the view, multiple frames of the inclined focus sensor are shown superimposed on the FOV 604 image sensor frame in intermediate positions at times 0, 1, 2 and 3. Focus points can be taken at three points between the centers of adjacent frames in the focus column. Focus positions 0, 1, 2, and 3 are used to extrapolate the best focus position to the next frame photographed at position 0 *. The scan time for this method should be similar to the methods described elsewhere here. While the FOV 602 frame of the inclined focus sensor has a shorter advance reading, in this case 0.707 x (0.94-0.432) / 2 = 0.18 mm or the inclined focus sensor invades 42% in the next frame to be acquired, the FOV frame 602 of the inclined focus sensor, which is oblique with respect to the FOV 604 image sensor frame, sees the fabric at the edges of the scan strip which may be advantageous in certain cases to provide edge focus information.
[0110] Figure 16 is a schematic illustration 650 showing an alternative arrangement for processing the focus according to another embodiment of the system described here. As in illustration 650, the FOV 652 frame of the tilted focus sensor and the FOV 654 frame of the image sensor are shown. The FOV 652 frame of the inclined sensor can be used to acquire focus information on the step forward through the fabric.
[0111] No pass back, the imaging sensor shoots frames while the focus stage adjusts using previous step forward focus data. If you want to take focus data at all intermediate image frame jump positions 0, 1, 2, 3 in the previous method, the Stage with XY movement can move 4x the speed in step forward given the high rate of point acquisition of focus. For example, for 15mm x 15mm to 20x, a column of data is 35 frames. Since the focus data is acquired at 120 points per second, the step forward can be performed in 0.3 seconds (35 frames / 120 focus points per second). The column number in this example is 26, so the focus portion can be done in 26 x 0.3 or 7.6 seconds. Image acquisition at 30 fps is approximately 32 seconds. Thus, the focus portion of the total scan time is only 20%, which is efficient. Furthermore, if they are allowed to skip any other frame, the focus portion of the scan time would fall even more substantially.
[0112] It has been found that in other embodiments, the focus sensor's focus strip can be positioned elsewhere within the field of view and in other orientations for sampling adjacent columns of data to provide additional read-ahead information that can be used in connection with the system described here.
[0113] The Stage with XY movement that carries the blade can repeat the points of better focus produced in the forward path in relation to those produced in the back path. For a 20x 0.75 NA lens where the depth of focus is 0.9 micron, it would be desirable to repeat up to approximately 0.1 micron. Internships can be built that meet 0.1 micron forward / backward repeatability and correspondingly this requirement is technically feasible as elsewhere discussed here.
[0114] In one embodiment, a fabric or stain on a glass slide being examined according to the system described here can cover the entire slide or approximately an area of 25 mm x 50 mm. Resolutions are dependent on the numerical aperture (NA) of the objective, the coupling means to the blade, the NA of the condenser and the wavelength of light. For example, at 60x, for a 0.9 NA microscope objective, flat apochromatic (Plan APO), in the air in a green light (532 nm), the lateral resolution of the microscope is approximately 0.2 μm with a depth of focus of 0.5 um .
[0115] In connection with the system operations described here, digital images can be obtained by moving a limited field of view via an in-line scanning sensor or CCD array over the area of interest and grouping the limited field of view or frames or slopes together to form a mosaic. It is desirable that the mosaic looks seamless with invisible suture, focus anomalies or irradiance when the viewer navigates the entire image.
[0116] Figure 17 is a flowchart 700 showing the processing to acquire a mosaic image of fabric on a slide according to an embodiment of the system described here. In a step 702, a thumbnail image of the slide can be acquired. The thumbnail image can be a low resolution in the order of a magnification of 1x or 2x. If a bar code is present on the slide label, the bar code can be decoded and attached to the slide image in this step. After step 702, processing takes place until step 704 where the fabric can be found on the slide using standard image processing tools. The fabric can be bonded to limit the scanning region to a certain area of interest. After step 704, processing takes place up to step 706 where an XY coordinate system can be joined to a fabric plane. After step 706, processing can occur up to step 708 where one or more focus points can be generated in regular X and Y spacing for the fabric and the best focus can be determined using a focus technique such as one or more real-time focusing techniques covered here elsewhere. After step 708, processing can take place up to step 710 where the coordinates of the desired focus points, and / or other appropriate information, can be saved and can be called anchor points. It was found that where frames are located between the anchor points, a focus point can be interpolated.
[0117] After step 710, processing can occur up to step 712 where the microscope objective is positioned in the best focus position according to the techniques discussed here at another point. After step 712, processing occurs until step 714 where an image is collected. After step 714, processing takes place until a test step 716 where it is determined whether an entire area of interest has been scanned and imaged. If not, the processing will take place until a step 718 where the XStage Y moves the fabric in the X direction and / or Y direction according to the techniques discussed here at another point. After step 718, processing takes place back to step 708. If in test step 716, it is determined that an entire area of interest has been scanned and imaged, processing will occur until step 720 where the collected frame images are stitched or otherwise combined to create the mosaic image according to the system described here and using techniques discussed here in another point (for example, with reference to the patent document US Patent App. Pub. No. 2008/0240613). After step 720, processing is complete. It has been found that other appropriate sequences can also be used in connection with the system described here to acquire one or more mosaic images.
[0118] For operation of the system described here advantageously, repeatability of z position can be repeatable up to a fraction of the depth of focus of the objective. A small error in the return to the z position by the focus motor is easily seen in a side-by-side system (2D CCD or CMOS) and in the adjacent columns of an in-line scanning system. For the above resolutions at 60x, a peak repeatability of z in the order of 150 nanometers or less is desirable and such repeatability should be correspondingly suitable for other objectives, such as 4x, 20x and / or 40x objectives.
[0119] According to the system described here, several embodiments for a slide stage system including an X Stage Y are provided for the application of pathology microscopy that can be used in connection with the digital pathology imaging features and techniques that are addressed here, including, for example, functioning as a Stage with XY 130 movement discussed here elsewhere in connection with real-time focusing techniques. According to one embodiment, and as discussed elsewhere here below, an X Stage Y may include a rigid base block. The base block may include a flat glass block supported on high projections and a second glass block with a triangular cross section supported on high projections. The two blocks can be used as smooth, straight rails or tracks to guide a moving stage block.
[0120] Figure 18 is a schematic illustration showing an implementation of a precision stage 800 (for example, a portion of Stage Y) of an X Stage Y according to an embodiment of the system described here. For example, precision stage 800 can achieve peak z repeatability in the order of 150 nanometers or less over an area of 25 mm x 50 mm. As discussed below, the precision stage 800 can be used in connection with features and techniques discussed here at another point, including, for example, operation in connection with the XY 130 movement stage presented with respect to focusing techniques. In real time. The precision stage 800 may include a rigid base block 810 where a flat glass block 812 is supported on high projections. The spacing of these saliencies is such that the bending due to the weight of the precision stage 800, the glass blocks on the simple supports are minimized. A second glass block 814 with a triangular cross-section is supported on high projections. The glass blocks 812, 814 can be adhesively bonded to the base block 810 with a semi-rigid epoxy that does not deform the glass blocks. The glass blocks 812, 814 can be straight and polished to one or two waves of light at 500 nm. A low thermal expansion material, such as Zerodur, can be employed as a material for glass blocks 812, 814. Other suitable types of glass can also be used in connection with the system described herein. An 816 cutout can allow light from a microscope condenser to illuminate the tissue on the slide.
[0121] The two glass blocks 812, 814 can be used as smooth, straight rails or tracks to guide the 820 moving stage block. The 820 moving stage block can include spherically shaped buttons made of plastic material (for example , 5 buttons) that contact the glass blocks, as shown in positions 821a-e. due to the fact that these plastic buttons are spherical, the contact surface can be confined to a very small area (0.5 mm) determined by the plastic's modulus of elasticity. For example, PTFE or another therroplastic mixture plus other lubricating additives manufactured by GGB Bearing Technology Company, UK can be used and fuse into the shape of the contact buttons approximately 3 mm in diameter. In one embodiment, the coefficient of friction between the plastic button and polished glass should be as low as possible but the use of a liquid lubricant should be avoided to save on instrument maintenance. In one embodiment, a coefficient of friction between 0.1 and 0.15 can be obtained printily in dry operation.
[0122] Figures 19A and 19B are more detailed views of the moving stage block 820 according to an embodiment of the system described here, showing the spherically shaped buttons 822a-e that contact the glass blocks 810, 812 in positions 821a-e. The buttons can be arranged in positions that allow excellent rigidity in all directions other than the drive direction (Y). For example, two plastic buttons can collate to contact sides of the 814 triangular shaped glass block (ie, 4 buttons 822b-e) and a plastic button 822a is positioned to contact the flat glass block 812. The stage block with Movement 820 may include one or more holes 824 to be reduced in weight and shaped to place the center of gravity in the centroid 826 of the triangle formed by the position of plastic support buttons 822a-e. In this way, each of the plastic buttons 822a-e at the corners of the triangle 828 can have the same weight at all times when moving a stage 800.
[0123] With reference to figure 18, a blade 801 is fixed via a spring-loaded arm 830 in the bundle 832. Blade 801 can be manually placed in bundle 832 and / or robotically placed in bundle 832 with an auxiliary mechanism. A rigid cantilever arm 840 supports and rigidly secures the end of the small-diameter rod flexes 842 that can be made of high-fatigue-resistant steel. In one example, this diameter can be 0.7 mm. The other end of the flexural rod 842 can be attached to the centroid location 826 in the movement stage 820. The cantilever arm 840 can be attached to a support block 850 which can operate through a recirculating support configuration on a tempered steel rail 852. A set of lead screw 854 can be attached to the support block 850 and the set of lead screw 854 can be rotated by a stepper motor 856. Components suitable for the above elements can be made available through various companies such as as THK in Japan. The lead screw set 854 drives the support block 850 over the rail 852 which pulls or pushes the stage block with movement 820 via the flexure of stem 842.
[0124] The flexural stiffness of the 842 rod flexure can be a factor greater than 6000x less than the stiffness of the 820 movement stage block in its plastic cushions (this is a stiffness opposite to a force orthogonal to the plane of the staging stage. movement in the Z direction). This effectively isolates the stage block with movement 820 against up and down movements of the support block 850 / cantilever arm 840 produced by the rolling noise.
[0125] Careful mass balancing and attention to geometry in the configuration of the precision stage 800 described here minimizes moments in the stage block with movement 820 that can produce small oscillating movements. In addition, since the 820 movement stage block is made of polished glass, the 820 movement stage block presents repeatability of the z position below the 150 nanometer peak sufficient for scanning with 60x magnification. As the 60x condition is the most stringent, other minor magnifications such as 20x and 40x high NA objectives also show adequate performance similar to the performance obtained under 60x conditions.
[0126] Figure 20 shows an implementation of an entire XY composite Stage 900 according to the precision stage characteristics discussed here and including a Y Stage 920, an X Stage 940 and a base plate 960 according to one embodiment of the system described here. In this case, a base block for Stage Y 920 becomes Stage X 940 which is a movement stage in the X direction. A base block for Stage X 940 is the base plate 960 that can be grounded. The composite stage XY 900 provides repeatability in the Z direction in the order of 150 nanometers and repeatability in the order of 1-2 microns (or less) in the X direction and Y direction according to the system described here. If the stages include a return position via a tape scale, such as that produced by Gloucestershire, England, sub-micron prices can be obtained according to the system described here.
[0127] The stage configuration according to the system described here can be superior to the movement stages supported by a spherical bearing where an X Stage Y according to the system described here does not suffer repeatability errors due to non-spherical ball bearings or non-cylindrical roller bearings. In addition, in recirculating bearing configurations, a new ball component with spheres of different sizes can cause non-repeatable movement. An additional benefit of the achievements described here is the cost of the internship. The glass elements use standard grinding and polishing techniques and are not very expensive. The bearing block and lead screw assembly do not need to be of particularly high quality in that the rod flexure decouples the movement stage of the bearing block.
[0128] According to the system described here, it is advantageous to reduce and / or minimize scan times during the digital scanning of pathology slides. In clinical configurations, a desired workflow is to place a slide holder on a robotic slide scanning microscope, close the door and command the system to scan the slides. It is desired that no user intervention is necessary until all the blades are swept. The batch size can include multiple slides (for example, 160 slides) and the time to sweep all slides is called batch time. Blade yield is the number of blades per hour processed. The cycle time is the time between each available slide image that is ready for viewing.
[0129] The cycle time can be influenced by the following steps in the acquisition of an image: (a) robotically grab the blade; (b) create a miniature or panoramic image of the tissue area for the label slide; (c) calculating an area of interest that connects a tissue to the slide; (d) pre-scanning the connected tissue area to find a regular set of points of better focus on the tissue; (e) sweeping the fabric according to the movement of a stage and / or sensor; (f) create a compressed output image ready for viewing; and (g) deposit the slide ready for the next slide. It was found that step (d) may not be necessary if dynamic or “real-time” focusing is performed according to the system described here, and in which image / scan time can be correspondingly reduced as a result using real-time focusing techniques.
[0130] The system described here can also include the elimination or significant reduction of the time for the execution of steps (a), (b), (c) and (g). According to several embodiments of the system described here, these gains can be realized, for example, by using a concept of caching where the steps mentioned above (a), (b), (c) and (g) for a blade are superimposed in time by steps (d), (e) and (f) for another blade as presented here at another point. In several embodiments, the overlapping of steps (a), (b) and (c) for a blade with steps (d), (e) and (f) for another blade can provide a gain of 10%, 25% or even even 50% in coparação with a system where steps (a), (b) and (c) for one slide are not overlapped by steps (d), (e) and (f) for another slide.
[0131] Figure 21 is a schematic illustration showing a caching device 1000 according to an embodiment of the system described here. A blade grab head 1002 can be positioned to pick up a blade 1001. The grab head 1002 can use a mechanical device and / or a vacuum device to pick up blade 1001. Blade 1001 can be one of the blade collection in the batch, for example, a batch of 160 blades. The blade collection can be arranged in a blade holder 1003 blade holder. The gripping head 1002 is attached to a cart or bearing block 1004 that moves over a steel rail 1005. The bearing block 1004 is moved by a rotating lead screw 1006. Motor counts can be detected with a rotary encoder 1007 and converted into a linear path to control the blade position in the Y direction. Elements 10021007 can comprise a movement set called a feeder / unloader blade loader / unloader 1008. Blade loader / unloader 1008 can also move a motorized bearing cart or motorized bearing block 1009 in the X direction on rail 1010 which allows the blade loader / unloader 1008 to move on both the X and Y directions.
[0132] During operation, a slide, while attached to the gripping head 1002, can be positioned under a low resolution camera 1011 to obtain the thumbnail image or panoramic image of the tissue area for the slide and label (for example, the step (b) above). Once this operation is completed, step (c) can be performed and the blade will be placed in a position on a 1012 blade temporary storage. The 1012 blade temporary storage can include two (or more) temporary storage slots or positions 1018a, 1018b, and is shown including a blade 1017 in the temporary storage position 1018a.
[0133] In one embodiment, a compound X13 Stage Y 1013 may include a 1014 stage plate that moves in the Y Direction and that is mounted on a 1015 plate that moves in the X direction. The X13 Stage Y 1013 may have similar features and functionality to that discussed here elsewhere, including, for example, characteristics of the XEstágio Y 900 compsoto discussed here. The stage plate 1014 may also include an adicoinal blade grip head 1016. The grip head 1016 may be similar to the grip head 1002 described above. The grip head 1016 can use a mechanical device and / or a vacuum device to pick up a blade.
[0134] The X16 Stage Y compound 1016 grip head 1013 can move to the temporary storage position 1018a and pick up the blade 1017. The blade 1017 can now proceed to one or more of the above steps, including the steps of: (d ) pre-scan, (e) scan and f) create output image steps. While this processing is being carried out, the blade loader / unloader 1008 can pick up another blade (for example, blade 1001), obtain the thumbnail image of blade 1001 using camera 1011, and place blade 1001 in an empty position 1018b on temporary storage for blade 1012, shown schematically by dotted line 1001 '. When sweeping is completed on the previous blade (blade 1017), the XY 1013 composite stage 1016 blade grab head can place blade 1017 in temporary storage position 1018a and pick up the next blade (blade 1001) from the temporary storage 1018b that is ready for scanning. The X13 Stage Y compound 1013 can move in a regular scanning pattern back and forth under high resolution optical microscope lenses and 1019 camera to acquire a high-resolution image of biological tissue according to characteristics and techniques here addressed at another point. It was also found that blade movements and selections of the compound XStage Y 1013 and / or blade loader / unloader 1008 can be controlled by one or more processors in a control system.
[0135] Blade loader / unloader 1008 can move to temporary storage position 1018a and pick up blade 1017 and deposit blade 1017 in slide holder 1003. This blade 1017 has completed all of the steps listed above. The blade loader / unloader 1008 can then continue to pick up and load another blade into the temporary storage for blade 1012, and if necessary pick up and return blade 1001 to the slide holder 1003. Processing like the above can continue until all slides that are in the 1003 slide holder have been swept.
[0136] Blade caching techniques according to the system described here provide advantageous time savings. For example, in a 20x 15 mm x 15 mm field system, the hold time is approximately 25 seconds, the thumbnail image acquisition is approximately 10 seconds, the preliminary scan time is approximately 30 seconds and the scan time is 90 seconds. The output file is generated simultaneously with the scanning process and can add approximately 5 seconds. The deposit of the blade is approximately 20 seconds. The addition of these times indicates a cycle time of 180 seconds. The Composite Stage XY still needs time to pick up and deposit the swept blade which can be approximately 10 seconds. Correspondingly, the scan time reduction is therefore approximately 1- (180-55 + 10) / 180 = 25%. For systems using dynamic focusing techniques such as real-time focusing as discussed below, the preliminary scan time needs to be eliminated and with high data rate cameras, times not associated with grab and deposit can be reduced to 20 -30 seconds. The reduction in scanning time using blade caching in this case can be approximately 1- (75-55 + 10) / 75 = 50%.
[0137] Figure 22A is a flowchart 1100 showing caching processing according to an embodiment of the system described here in connection with a first slide. In a step 1102, the first blade is picked up from a blade holder. After step 1102, processing takes place until step 1104 where a miniature image is obtained and / or other miniature processing, which may include determining an area of tissue interest on the slide is performed for the first slide. After step 1104, processing takes place until step 1106 where the first blade is deposited in a temporary blade storage. After step 1106, processing takes place until step 1108 where the first blade is picked up from the blade buffer. After step 1108, processing occurs until step 1110 where the first blade is scanned and imaged according to techniques such as the one discussed here elsewhere. It was found that in various embodiments the scanning and imaging techniques may include pre-scan focusing steps and / or the use of dynamic focusing techniques, such as a real-time focusing technique. After step 1110, processing takes place until step 1112 where the first blade is deposited in the temporary blade storage. After step 1112, processing takes place up to step 1114 where the first blade is picked up from the blade buffer. After step 1114, processing takes place until step 1116 where the first slide is deposited on the slide holder. After step 1116, processing is complete with respect to the first blade. 22B is a flow chart 1120 showing caching processing according to an embodiment of the system described herein in connection with a second blade. As discussed here, several steps of flowchart 1120 can be performed in parallel to the steps of flowchart 1100. In one step 1122, the second blade is picked up from a slide holder. After step 1102, processing takes place until step 1124 where a miniature image is obtained and / or other processing for miniature, which may include determining an area of tissue interest on the slide, is performed for the second slide. After step 1124, processing takes place until step 1126 where the second blade is deposited in a temporary blade storage. After step 1126, processing takes place up to step 1128 where the second blade is picked up from the blade buffer. After step 1128, processing takes place until step 1130, where the second blade is scanned and imaged according to techniques such as the one discussed here elsewhere. It was found that in various embodiments the scanning and imaging techniques can include pre-scan focusing steps and / or the use of dynamic focusing techniques, such as real-time focusing technique. After step 1130, processing takes place until step 1132, where the second blade is deposited in the temporary blade storage. After step 1132, processing takes place until step 1134 where the second blade is picked up from the temporary blade storage. After step 1134, processing takes place until step 1136 where the second blade is deposited on the slide holder. After step 1136, processing is complete with respect to the second blade.
[0138] According to an embodiment of the system described here, blade caching addressing, steps of flowchart 1100 with respect to the first blade can be performed via a caching device parallel to the steps of flowchart 1120 with respect to the second blade to reduce cycle time. For example, steps 1122, 1124, 1126 of flowchart 1120 for the second slide (for example, the steps in connection with grabbing the second slide of the slide holder, miniature image processing and depositing the second slide in the slide buffer ) can be overlapped with steps 1108, 1110, and 1112 of flowchart 1100 with respect to the first slide (for example, the steps in connection with grabbing the first blade of the temporary storage for slide, scanning and imaging the first slide and depositing the first slide blade back to the blade temporary storage). In addition, steps 1134 and 1136 (for example, steps in connection with grabbing the second blade of the blade buffer and depositing the blade in the blade holder) can also overlap with the scanning steps of the first blade. Time gains of up to 50% can be obtained according to the parallel blade processing techniques according to the system described here in comparison to the processing of a blade in one time, with additional gains possible using other aspects of the system and techniques described here.
[0139] Figures 23A and 23B show time diagrams using blade caching techniques in accordance with the embodiments of the system described herein and illustrate time savings in accordance with various embodiments of the system described herein.
[0140] Figure 23A shows time diagram 1150 for the scenario in which a pre-scan step is used. The time diagram shows the synchronization for three slides (Blades 1, 2 and 3) during an interval of approximately 300 seconds in connection with the execution of the blade processing steps using blade caching including grabbing a blade from a slide holder, miniature image processing, slide deposition in the temporary storage, grab from the temporary storage, pre-scan, slide scanning and file output, deposition in the temporary storage and deposition in the slide holder. As illustrated, in one embodiment, the cycle time for the illustrated processing can be approximately 150 seconds.
[0141] Figure 23B shows time diagram 1160 for a scenario in which a real-time focusing technique is used (without pre-scanning). The time diagram shows the synchronization for three slides (Blades 1, 2 and 3) during an interval of approximately 150 seconds in connection with the execution of the blade movement and sweeping steps using blade caching including grabbing a blade. blade from a slide holder, miniature image processing, slide deposition in the temporary storage, grab from the temporary storage, slide scanning and file output, deposition in the temporary storage and deposition in the slide holder. As illustrated, in one embodiment, the cycle time for the illustrated processing can be approximately 50 seconds.
[0142] Figure 24 is a schematic illustration showing a caching device 1200 according to another embodiment of the system described here. In the illustrated embodiment, no temporary storage is required, and grab, miniature and deposit times can be eliminated from cycle time using the caching device 1200. The caching device 1200 can include two compound stages XY 1210 , 1220 that operate independently. Each of the composite stages XY 1210, 1220 can have characteristics similar to those discussed here with respect to the composite stage XY 1013. A first support for slides 1211 can be positioned at one end of stage 1210 and a second support for slides 1221 can be positioned at one end of stage 1220. It has been found that in connection with another embodiment of the system described herein, the first blade holder 1211 and the second blade holder 1211 can refer to portions of a blade holder. Two miniature cameras 1212, 1222 can serve each of the composite stages XY 1210, 1220. Each of the slide holders 1211, 1221 can serve slides and their companion composite stage XY 1210, 1220 with a corresponding grip head. An optical microscope train 1230 can serve both compound stages XY 1210, 1220. For example, while one of the compound stages XY (for example, stage 1210) is sweeping a slide, the other (for example, stage 1220) is performing the operation to pick it up, the thumbnail and deposit functions with another slide. These functions can be overlapped with the scan time. Correspondingly, the cycle time can be determined by the scanning time of a slide and holding times, miniature image and deposit are therefore eliminated from the cycle time according to the illustrated embodiment of the system described here.
[0143] Figure 25A is a flowchart 1250 showing caching processing in connection with a first blade according to an embodiment of the described system for a caching device featuring two composite XY stages for blade processing. In a 1252 step, the first blade is picked up from a blade holder. After step 1252, processing takes place up to step 1254 where processing for miniature is performed on the first slide. After step 1254, processing takes place until step 1256 where the first slide is scanned and imaged according to techniques like the one discussed here elsewhere. It was found that in various embodiments the scanning and imaging techniques may include pre-scan focusing steps and / or the use of dynamic focusing techniques, such as a real-time focusing technique. After step 1256, processing takes place until step 1258 where the first blade is deposited back in the slide holder. After step 1258, processing is complete with respect to the first slide.
[0144] Figure 25B is a flowchart 1270 showing caching processing in connection with a second blade according to an embodiment of the system described for a caching device with two composite XY Stages for blade processing. In a 1272 step, the second blade is picked up from a blade holder. After step 1272, processing takes place until step 1274 where processing for miniature image is performed on the second slide. After step 1274, processing takes place until step 1276 where the second blade is scanned and imaged according to techniques such as the one discussed here at another point. It was found that in various embodiments the scanning and imaging techniques may include pre-scan focusing steps and / or the use of dynamic focusing techniques, such as a real-time focusing technique. After step 1276, processing takes place until step 1278 where the second blade is deposited back in the slide holder. After step 1278, processing is complete with respect to the second blade.
[0145] According to an embodiment of the system described here involving blade caching, steps in flowchart 1250 with respect to the first blade can be performed by the caching device parallel to the steps in flowchart 1270 concerning the second blade in order to reduce cycle time. For example, steps 1272, 1274 and 1278 for the second slide (for example, grab, thumbnail and deposit processing) can override step 1256 for the first slide (for example, scan / scan the first slide), and vice vice versa, so that grab times, thumbnail image processing and deposit times are eliminated from cycle time. The cycle time is correspondingly determined by only the scanning time of a slide according to an embodiment of the system described here.
[0146] Figure 26 is a schematic illustration showing a 1300 caching device according to another embodiment of the system described here. The 1300 caching device can include a blade holder configured as a 1310 carousel, a 1320 blade handler, a 1330 buffer and an XY 1340 Stage. The 1310 carousel can include one or more blade holder devices (for example , cassettes) that define positions 1312, 1312 ', 1312' 'in which slides, such as blade 1301, can be placed before and / or after being imaged by a 1350 imaging device that can present features and functionality like the one discussed here in other point.
[0147] Positions 1312, 1312 ', 1312' 'are shown as a set of wedges (for example, 8 wedges) and, as discussed below, carousel 1310 can have a height so that multiple blade positions extend below each of the top level wedge positions 1312, 1312 ', 1312' 'that are shown. The blade handler 1320 may include a arm 1322 that acts as a gripping head and may include mechanical and / or vacuum devices for picking up a blade. The arm 1322 on a blade manipulator 1320 can move between positions 1322a-d to move blades between carousel 1310, temporary storage 1330 and Stage XY 1340.
[0148] The temporary storage 1330 can include multiple temporary storage positions 1332, 1334. A temporary storage position 1332 can be designated as a return position of the temporary storage 1332 in which slides that are returning from the imaging device 1350 via Stage XY 1340 can be positioned before being moved by the blade handler 1320, back to the carousel 1310. Another 1334 staging position can be designated as a 1334 camera staging position, in which a blade that must be sent to the device of imaging 1350 can first present a miniature image captured from the slide according to the techniques discussed here at another point. After a miniature image of the slide is captured in the camera temporary storage position 1334, the slide can be moved to a position 1342 in Stage XY 1340 that transports the slide to the imaging device 1350 for scanning and imaging according to techniques here covered in another point.
[0149] Figure 27 is a schematic illustration showing another view of the 1300 caching device. The components of the 1300 caching device may have functionality to operate with multiple movements and with multiple degrees of freedom of movement. For example, carousel 1310 can be rotated in one direction 1311 and can include multiple blade positions 1312a-d at multiple height positions in each rotational position to accommodate multiple blades (shown as Blades 1, 2, 3 and 4). In one embodiment, the multiple blade positions 1312a-d in each of the wedge positions 1312, 1312 ', 1312' 'can include positions for 40 blades, for example, positioned equidistantly within the height of the carousel 1310 that you can measure in one embodiment, 12 inches. In addition, carousel 1310 may also include a user tray 1314 with one or more blade positions 1314a, b in which a user can insert a blade to be imaged in addition to other slides in the 1310 carousel. Interaction of a blade in the tray user 1314, for example lifting a cover of user tray 1314 and / or inserting the blade at one end of positions 1314a, b of user tray 1314, can act to trigger a bypass mode in which a blade of the user tray 1314 is processed instead of the next blade from the wedge positions of the 1310 carousel.
[0150] The arm 1322 of the blade manipulator 1320 is shown with at least three degrees of freedom of movement. For example, the arm 1322 can rotate in a direction 1321a in order to engage each carousel 1310, the temporary storage 1330 and the Stage XY 1340. Additionally, the arm 1322 can be adjustable in a direction 1321b corresponding to different heights of positions 1312a- d of carousel 1310. Additionally, the arm 1322 can extend in direction 1321c in connection with loading and unloading blades of carousel 1310, temporary storage 1330 and Stage XY 1340. In one embodiment, it is advantageous to minimize the arc distance that the arm 1322 rotates and / or minimizes other distances crossed by the arm 1322 and / or blade manipulator 1320 in order to minimize dead times of the caching device 1300, as shown below. Movements of carousel 1310, blade manipulator 1320, and Stage XY 1340 can be controlled in several embodiments by a control system like the one discussed here elsewhere. It was also found that, in one embodiment, the temporary storage 1330 and Stage XY 1340 can be at the same level.
[0151] The slides can carry different types of biological samples. A biological sample can be a tissue sample (for example, any collection of cells) removed from an object plant. In some embodiments, a biological sample includes, without limitation, a tissue section, an organ, a tumor section, a spot, a frozen section, a cytology prep, or cell lines. An incisional biopsy, a fragment biopsy, an excisional biopsy, a needle aspiration biopsy, a fragment needle biopsy, a stereotactic biopsy, an open biopsy, or a surgical biopsy can be used to obtain the sample. In some embodiments, the sample may be a section of a plant, plant tissue culture or the like.
[0152] Slides can generally be flat transparent substrates that can carry samples for examination using equipment, such as optical equipment, for example, a microscope or other optical device. For example, blade 1301 of figure 26 can be a generally rectangular piece of transparent material with a front face for supporting a sample. The microscope slide 1301 can be in the form of a standard microscope slide made of glass or other transparent material. In some embodiments, the blade 1301 is approximately 3 inches (75 mm) long, approximately 1 inch (25 mm) wide and approximately 1 mm thick.
[0153] Blade 1301 may include a label. Labels may include machine-readable code (such as a mono- or multidimensional bar code or infoglyph, an RFID tag, a Bragg diffraction grid, a magnetic strip or a nanobar code) with coded instructions (for example, instructions for imaging), object information, tracking information or the like. The tag can be scanned by readers located at different locations on the 1300 caching device.
[0154] The slide may include coverslips to protect the samples. If blade 1301 is a standard microscope slide, a coverslip may have a range in the range of approximately 0.5 inches (13 mm) to approximately 3 inches (76 mm), a width in the range of approximately 0.5 inch (13 mm) to approximately 1 inch (25.5 mm), and a thickness in a range of approximately 0.02 inches (0.5 mm) to approximately 0.08 inches (2 mm). In some embodiments, standard sipes with a length of approximately 50 mm, a width of approximately 24 mm, and a thickness of approximately 0.2 mm are used. Other dimensions are also possible, if necessary or desired. Lamellae can be made, entirely or only partially, from one or more polymers, plastics, composites, glass, combinations of these or other suitable materials that can in general be rigid, semi-rigid or deformable.
[0155] Figures 28A-28J are schematic illustrations showing blade caching operations of the caching device of figures 26 and 27 according to an embodiment of the system described herein. According to one embodiment, the blade operations discussed herein minimize system dead times, that is, the times during blade grab and transfer operations that do not overlap with blade scanning and imaging operations. Dead times can include, for example, a parking time where Stage XY 1340 moves into position to allow the 1320 blade handler to pick up the blade. Other contributions to dead time include moving the blade to the return position of the temporary storage 1330 and refilling the Stage XY 1340 with a blade.
[0156] Figure 28A starts the illustrated sequence in which a slide 2 is currently being scanned and imaged on the imaging device 1350. Blades 1, 3 and 4 are waiting to be scanned and imaged on the 1310 carousel, and the blade manipulator 1320 is in position to supply Blade 2 to Stage XY 1340. Figure 28B shows that blade manipulator 1320 rotates and descends to load the next blade (blade 3) to be scanned and imaged, while blade 2 continues to be swept and imagined. Figure 28C shows that the blade manipulator 1320 transports blade 3 to the camera temporary storage 1334 position of the temporary storage 1330 so that the thumbnail image is obtained from the blade 3. Figure 28D shows that the blade manipulator 1320 is positioned to unload the slide 2 of Stage XY 1340 that is, it returns from the imaging device 1350 after scanning of slide 2 has been completed. It has been found that the time when Stage XY 1340 moves to the position to be discharged is an example of time off. The time after Stage XY 1340 is in a position to be unloaded with blade 2 waiting there to be unloaded and blade 3 waiting to be loaded at Stage XY 1340 is an example of parking time.
[0157] Figure 28E shows that blade 2 is transported by blade manipulator 1320 from Stage XY 1340 to return position 1332 of buffer 1330. Blade manipulator 1320 then continues to the position to pick up blade 3 from from the camera temporary storage position 1334. Figure 28F shows that the blade 3 is picked up from the camera temporary storage position 1334 and discharged in Stage XY 1340. Figure 28G shows that the blade 3 is currently being swept, while the blade 2 is being picked up from the temporary storage return position 1332 by the blade manipulator 1320. Figure 28H shows that blade 2 is returned to its position on carousel 1310 by the blade manipulator 1320 which rotates and translates translationally to the appropriate position. Figure 28I shows that blade manipulator 1320 moves translationally to the proper position to pick up blade 1 from carousel 1310. Figure 28J shows that blade manipulator 1320 transports and unloads blade 1 in the temporary camera storage position where the thumbnail image of slide 1 is taken, while slide 3 is currently being scanned. Other iterations, similar to those discussed above in connection with the sequencing illustrated, can be performed against any remaining blade (for example, blade 4) on the 1310 carousel and / or for any user blade inserted by the user in the user tray 1314 for start the bypass mode discussed here.
[0158] Also according to the system described here, a lighting system can be used in connection with microscopy embodiments that are applicable to various techniques and characteristics of the system described here. It is known that microscopes in general can use Kohler illumination for white field microscopy. Primary characteristics of Kohler lighting are those in which the numerical aperture and illumination area are both controllable via adjustable irises so that the illumination can be adapted to work a wide range of microscope objectives with variable magnification, field of view and numerical aperture. Kohler lighting offers desired results but may require multiple components that take up a significant amount of space. Correspondingly, various embodiments of the system described here also provide characteristics and advantageous lighting techniques in microscopy applications that avoid certain disadvantages of the well-known Kohler lighting system while preserving the Kohler lighting advantage.
[0159] Figure 29 is a schematic illustration showing a lighting system 1400 for lighting a blade 1401 using a light emitting diode (LED) lighting set 1402 according to an embodiment of the system described here. The 1402 LED lighting kit can have several characteristics according to multiple embodiments as discussed here. Light from the LED lighting kit 1402 is transmitted through a mirror 1404 and / or other suitable optical components to a capacitor 1406. The capacitor 1406 can be a capacitor with a suitable working distance (for example, at least 28 mm) to accommodate any required working distance of an XY 1408 Stage, as discussed below. In one embodiment, the capacitor can be SG03.0701 condenser manufactured by Motic with a working distance of 28 mm. The condenser 1406 may include an adjustable iris diaphragm that controls the numerical aperture (cone angle) of light that illuminates the specimen on a slide 1401. The slide 1401 can be arranged in Stage XY 1408 under a microscope objective 1410. The 1402 LED lighting kit can be used in connection with scanning and imaging the specimen on a 1401 slide, including, for example, operations in relation to the movement of an XY Stage, blade cache storage and / or dynamic focus, according to the characteristics and techniques of the system described here.
[0160] The LED lighting kit 1402 can include an LED 1420, such as a pure white LED white, a 1422 lens that can be used as a collecting element, and an adjustable iris field diaphragm 1424 that can control the area of illumination on blade 1401. The emitting surface of LED 1420 can be imaged through lens 1422 on an inlet pupil 1406a of condenser 1406. Inlet pupil 1406a can be placed with an NA adjustment diaphragm 1406b of condenser 1406. The lens 1422 can be selected to collect a large fraction of the 1420 LED output light and also to focus an image of LED 1420 on NA 1406b adjustment diaphragm of condenser 1406 with appropriate magnification so that the image of LED 1402 fills the aperture of the Adjustment diaphragm NA 1406b of capacitor 1406.
[0161] Condenser 1406 can be used to focus the LED light 1420 on blade 1401 with the NA 1506b adjustment diaphragm. The illumination area on the blade 1401 can be controlled by the field diaphragm 1424 mounted on the LED lighting kit 1402. The field diaphragm, and / or spacing between capacitor 1406 and field diaphragm 1424, can be adjusted to image the light from LED 1420 in the plane of blade 1401 so that field diaphragm 1424 can control the area of blade 1401 that is illuminated.
[0162] As an image sensor acquires frames while a Stage Y containing a slide is moving, LED 1420 can be pulsed on and off (eg strobed) to allow very high brightness for a short time. For example, for a Y Stage that moves at approximately 13 mm / second, to maintain a maximum darkness of 0.5 pixels (0.250 micron / pixel), LED 1420 can be pulsed for 10 microseconds. The LED light pulse can be triggered by a master clock locked to the lens resonant frequency with Dither pattern according to the focus system and techniques discussed here elsewhere.
[0163] Figure 30 is a schematic illustration showing a more detailed image of an embodiment of an LED lighting kit 1402 'according to the system described here and corresponding to the characteristics described here in relation to the LED lighting kit 1402. One implementation and configuration of an LED 1430, a lens 1432, and a field diaphragm 1434 are shown with respect to and in connection with other structural support and adjustment components 1436.
[0164] Figure 31 is a schematic illustration showing an exploded view of a specific implementation of a 1402 '' LED lighting kit according to an embodiment of the system described here with characteristics and functions equal to those presented in relation to lighting kit LED 1402. An adapter 1451, holder 1452, holder 1453, and holder 1454 can be used to safely mount and place an LED 1455 in the LED lighting kit 1402 '' in order to be positioned securely with respect to a 1462 lens. Appropriate components screw and washer 1456-1461 can also be used to attach and assemble the 1402 '' LED lighting kit. In various embodiments, LED 1455 can be a Luminus, PhlatLight White LED CM-360 Series this is a pure white LED with an optical output of 4,500 lumens and a long life of 70,000 hours and / or a suitable LED manufactured by Luxeon. A 1462 lens can be a MG 9P6mm lens, 12mm OD (outside diameter). A 1463 tube lens component, 1464 adapter, stack tube lens component and 1467 retaining ring can be used to position and mount a 1462 lens with respect to the 1465 adjustable field diaphragm component. The field diaphragm component adjustable 1465 can be a ring-activated iris diaphragm, part number SM1D12D from Thor Labs. The 1466 stack tube lens can be a Thor Labs P3LG lens. The 1463 tube lens can be a P50D or P5LG tube lens from Thor Labs. Other washer components 1468 and screw components 1469 may be used, if appropriate to secure and assemble elements of the 1402 '' LED lighting kit.
[0165] According to the system described here, devices and techniques are provided for high-speed blade scanning for digital pathology applications according to various embodiments of the system described here. In one embodiment, a slide holder for a pathology microscope can include: (i) a disc-shaped tray and (ii) a plurality of recesses formed in the tray, in which each recess is adapted to receive a slide and the recesses are arranged circumferentially in the tray. The tray may include a central shaft hole and two locking holes, the locking holes being adapted to grip a gear adapted to rotate at high speed about a normal axis in relation to the tray. The recesses can be machined recesses at different angular positions on the tray. The recesses have semi-circular protrusions to touch the blade but not to over-tighten the blade, thus allowing the blade to be substantially free of deformation. The recesses may also feature a cutout that allows a finger clamp to place and extract the blade from the recess through an operator. In various embodiments, the blade holder and its operation can be used in connection with the features and techniques discussed here at another point for an imaging system.
[0166] Figure 32 is a schematic illustration showing a high-speed blade scanner 1500 according to an embodiment of the system described here that can be used in connection with digital pathology imaging. A blade holder 1510 can include a tray 1512 with recesses 1514a, b ... n arranged at angular positions of a circumferential or annular ring 1515 on tray 1512, and the recesses 1514a-n can be sized respectively to hold a blade 1501. Tray 1512 is illustrated as a circular disc and can be manufactured to hold a desired number of blades. For example, to hold 16 blades, tray 1512 can measure approximately 13 inches in diameter. It has been found that other blade configurations and tray size and shape can be used, if necessary, in connection with the system described herein, and the orientation and configuration of recesses 1514a-n and can be appropriately modified. A blade can be placed in each recess 1514a-n of tray 1512, just like placing blade 1501 in recess 1514a, and tray 1512 can be placed on the high speed blade scanner 1500. Tray 1512 can include a central shaft hole 1516c and two locking holes 1516a and 1516b, which can engage with a gear that rotates blade holder 1510 at high speed around axis 1518 in rotational direction 1519. Tray 1512 can be placed in a flat drawer, shown as 1502, which can retract tray 1512 for device 1500.
[0167] Figure 33 is a schematic illustration showing a 1520 recess in a high speed blade scanner tray in more detail according to an embodiment of the system described herein. The recess 1520 can be any of the recesses 1514a-n. The recess 1520 may include a plurality of semi-circular protrusions, such as three protrusions 1522a-c, to touch the blade 1501 but not to over-tighten the blade 1501, thus allowing the blade 1501 to be substantially free of deformation. A cutout 1523 allows a donor clamp to place and extract blade 1501 from recess 1520 through an operator. Centriped accelerations, shown schematically by sets 1521, produced by blade holder 1510 / tray 1512 when rotating around axis 1518 can apply a small holding force to blade 1501 to hold blade 1501 in place while imaging occurs. The holding force can be configured to be at least 0.1 g’s initially by rotating tray 1512 at rates greater than 100 rpm to register blade 1501 against semi-circular protrusions 1522a-c. Once the blade 1501 is registered, the rotation rate can be reduced consistent with system imaging rates such as those discussed here elsewhere. At lower rates, even a slight holding force should stabilize blade 1501 against protrusions 1522a-c.
[0168] Again with reference to figure 32, a microscope imaging system 1530, such as the one discussed in detail elsewhere here, can be arranged above the rotating whiting 1512 to image areas of the circumferential ring 1515 where the slides are placed. The imaging system 1530 may include a microscope objective 1532 of high NA, for example 0.75 of NA with a long working distance, an intermediate lens 1534 and a CCD or CMOS 2D image sensor 1536 placed at the appropriate distance to magnify objects on a slide 1501 for the image sensor 1536. The image sensor 1536 can have a high frame rate, such as over 100 frames / second. For example, the 1536 image sensor may be part of a Dalsa Falcon 1.4M100 camera that operates at 100 frames / second or the equivalent. The 1530 imaging system can be rigidly mounted on a motorized 2-axis drive that can be built from components such as DC motors or stepper motors, ball or lead screws or linear guides. One axis, the radial axis 1531a can move the imaging system 1530, or at least one component of it, radially through small movements, for example 1 mm steps with a resolution of 10 microns to image one or more rings in the spinning tray 1512 below. The other axis, the 1531b focus axis, moves in small 5-10 micron movements with 0.1 micron resolution. The focus axis can be built to perform movements at high speed, for example by executing a small movement in a few milliseconds. Movement of the 1534 microscope objective can be controlled through a control system and can be used in connection with dynamic focusing techniques such as those discussed here elsewhere.
[0169] A lighting system 1540 can be placed below the revolving tray 1502 and includes a light source 1542, such as a high-gloss white ED, one or more optical path components such as a mirror 1544, and a condenser 1546, similar to lighting components discussed here elsewhere. In one embodiment, the microscope condenser and imaging routes can be connected together and move as a rigid body such as a direction 1541 of movement of the lighting system 1540 is in the same direction as in the radial direction 1531a of the imaging system 1530. In the focus direction 1531b, the imaging path can be decoupled from the condenser path, such that one or more components of the imaging system 1530 can include independent movement in the focus direction 1531b to perform high-speed focus movements.
[0170] Figure 34 is a schematic illustration showing an imaging route that starts in a first radial position with respect to blade 1501 for imaging a specimen 1501 'on blade 1501 in recess 1520. Recess 1520 with blade 1501 rotates with the blade holder 1510 in the rotational direction 1524. Images can be captured for frames (for example, frames 1525) according to the image capture techniques discussed here at another point. As shown, images are captured for a row of frames (for example, frames 1525) for each slide on a slide holder 1510 when tray 1512 rotates under the imaging system 1530. After a complete revolution of tray 1512, the radial position of the Imaging system 1530 is implemented to capture images for another row of frames for each slide. Each frame is acquired at a high rate by temporarily freezing the next scene. Bright field lighting can be sufficiently radiant to allow for such short exposures. These exposures can be in a time frame of a few 10’s to a few hundred microseconds. The process is continued until the entire area of interest for each blade in the 1510 blade holder is imaged. In connection with this embodiment, processing the images collected in a mosaic image of an area of interest requires adequate organization mechanisms and / or image labeling to correctly correlate the multiple rows of frames between the multiple slides that are rotated on the 1512 tray. Appropriate imaging process techniques can be used to mark images in order to correlate captured images with the appropriate slide, since the movement of the image collection divided into blocks can be addressed by known image splicing software and can be transformed into images that a pathologist understands while looking at it under a standard microscope.
[0171] As an example, with a 13.2-inch diameter disk-shaped tray that rotates at 6 rpm, a 20x NA microscope objective = 0.75 produces a field of view of approximately 1 mm square. This arched field of view is traversed in approximately 10 msec. For a section of tissue within a 15 mm square active area and assuming 25% overlap between fields, 20 fields need to be incremented along the radial axis. If the frame transfer is short enough to limit acquisition time, 20 complete revolutions should be sufficient to image 16 slides on the disk. This should occur at 6 rpm in 200 seconds or a 1 blade yield every 12.5 seconds.
[0172] Figures 35A and 35B are schematic illustrations showing an alternative arrangement of blades on a rotating blade holder according to another embodiment of the system described here. Figure 35A shows a tray 1512 'with recesses 1514' configured in such a way that the longest dimension of the blade 1501 is oriented along the radius of the tray 1512 'in the shape of a disk that rotates in the direction 1519'. In this configuration, more blades (for example, 30 blades) can be accommodated on tray 1512 '. Fig. 35B is a schematic view showing an imaging path for blade 1501 in a recess 1520 ', that is, configured as referred to above. In the illustrated embodiment, blade 1501 is held in recess 1520 'according to centripedal forces shown in direction 1521' and protrusions 1522a'-c '. The direction of rotation 1524 'in which the image processing is performed is shown for the image collection of frames 1525' for the specimen 1501 '. The radial position of the 1530 imaging system is increased by longitudinal increments of the slides to capture images for successive rows of frames for each slide. In one example, for an area of 15 mm x 15 mm and assuming 25% overlap between fields. Twenty fields need to be incremented along the radial axis. Again, 20 revolutions at 6 rpm provide complete imaging in 200 seconds but with more efficient sweeping given the blade orientation and therefore yield should increase for one blade every 6.67 seconds.
[0173] Figure 36 is a schematic illustration showing an imaging system 1550 according to an embodiment of the system described here that includes an objective 1552 arranged to examine a specimen 1551 'on a blade 1551. In one embodiment, positions of focus can be predetermined by slower disc rotation before image acquisition. Survey of 20 seconds per slide for autofocus should total scan time below 30 seconds per slide - an order of magnitude faster than the current position of the systems according to the state of the art. When a tray 1560, on which the blade 1551 is placed, rotates in the direction 1561, the objective 1552 can pass through minute movements in the direction 1562 to be positioned in the best focus determined according to the system described here. Different autofocus values do not need to be adjusted for each 1553 field of view but apply to different larger zones 1554 on a 1551 blade, for example 3 x 3 fields of view or subframes due to higher spatial frequencies of blade warping or thickness of fabric. The auto-focus values should be interpolated by applying the best focus while the blade moves under the camera in its arc path.
[0174] Alternatively, a dynamic focusing technique, such as real-time focusing techniques described elsewhere here, can be advantageously employed in connection with the high-speed scanning systems provided here. It was found that the times to acquire focus points (for example, 120 focus points per second) allow the use of real-time focusing along with the high-speed rotational scanning techniques discussed here. It has also been found to be well within the field of control systems to control a rotational disk at speeds within 1 part of 10,000, allowing for open loop sampling of each image without relying on the rotational disc return.
[0175] Generally, a low resolution miniature image was produced from the slide. This can be done by adjusting a low-resolution camera on an angular position of the disc so as not to interfere with the high-resolution microscope just described. For extremely high volume applications, the disk format lends itself to robotic handling. Semi-conductor wafer robots that handle 300 mm (~ 12 ") disks can be used to move disks from a temporary stock to the high-speed scanning device. In addition, most technologies position the blade under the microscope objective through linear stages in one step and repetition movement. These movements dominate the image acquisition times. The system described here that uses a rotational motion is efficient and highly repeatable. Autofocus and image acquisition times are one order of magnitude lower than the current state of the art products.
[0176] Large systems also require clamping mechanisms or spring fasteners to hold the blade in place during the stop and advance movements of the stage. The system described here does not require a clamping mechanism in which the rotational movement creates centripetal acceleration that pushes the blade to a predetermined location within a recess cutout in the disc. This makes the construction of the blade holder simpler and more reliable. In addition, blade fasteners can bend or deform the blade, complicating the autofocus process and are advantageously avoided according to the system described herein.
[0177] Current systems present peak speeds of 2-3 minutes for an active area of 15 mm per blade. The systems and methods provided here allow the same active area to be scanned in less than 30 seconds, for the example highlighted above. Many pathology laboratories sweep from 100 slides to 200 slides per day. With these high rates of image acquisition an operator could work with a daily stock of slides in an hour including the additional steps of loading and unloading discs, barcode reading, pre-focus. This allows faster time for results and greater savings for the laboratory.
[0178] Figure 37 is a flow chart 1600 showing high-speed blade scanning using a rotating tray according to an embodiment of the system described here. In a 1602 step, blades are located in recesses of the turntable. After step 1602, processing takes place up to step 1604 where the rotating tray is moved to a blade scan position with respect to scanning and imaging system. After step 1604, processing takes place until step 1606 where the rotation of the rotating tray is initiated. As discussed above, the rotation of the rotating tray produces centripedal forces that act on the blades to keep the blades in a desired imaging position. After step 1606, processing takes place until step 1608 where the imaging system captures images, according to the systems and techniques described here and including dynamic focusing techniques, for a row of frames for each slide in a circumferential ring of the rotating tray. After step 1608, processing takes place until a test step 1610 where it is determined whether an area of interest desired on each slide in the turntable has been scanned and imaged. If not, the processing takes place up to a step 1612 where the imaging system and / or certain components of it move an increment in a radial direction of the rotating tray. After step 1612, processing takes place back to step 1608. If in test step 1610, it is determined that the area of interest on each slide has been scanned and imaged, processing takes place until step 1614 where one or more mosaic images are created correspondingly to the areas of interest imagined for each slide. After step 1614, processing is complete.
[0179] According to the system described here, an optical and technical duplication device can be provided and used in connection with the characteristics of the imaging system described here. In one embodiment, the system described here can sample a resolution element produced by a 20x 0.75 NA Plan Apo objective. This resolution element is approximately 0.5 microns at a wavelength of 500 nm. To sample this resolution element, the tube lens in front of the imaging sensor can be changed. An approximate calculation for computing the focal length of the tube lens given the objective lens (tube lens = focal length of the tube lens in front of the image sensor) is: pix_sensor = pixel size in CCD or CMOS image sensor pix_object = pixel size in object or fabric tube lens = pix_object / pix_sensor * 9 mm.
[0180] To obtain an object pixel size of 0.25 micron for Dalsa Falcon 4M30 / 60 (7.4 micron sensor pixel), the focal length of the tube lens should be approximately 266 mm. For an object pixel size of 0.125 micron, the focal length of the tube lens should be approximately 532 mm. It may be desired to change the pixel sizes between these two objects and this can be done by fitting two or more tube lenses to a stage that moves in front of the imaging sensor. Given the different path lengths associated with each new focal length, fold mirrors also need to be added to bend the path of a fixed position image sensor.
[0181] Figure 38 is a schematic illustration showing a 1700 optical duplication image system according to an embodiment of the system described here. The 1700 optical duplication image system may include a 1710 image sensor from a 1711 camera and a 1720 microscope objective as described elsewhere here. It has been found that other components in connection with the system and techniques discussed here, such as real-time focusing system, can also be used with the illustrated 1700 optical duplication image system. To obtain two or more pixel sizes, a plurality of tube lenses, for example, a first tube lens 1740 and a second tube lens 1750, can be provided in connection with the system described herein. A stage 1730 can move the first tube lens 1740 and the second tube lens 1750, respectively, in front of the imaging sensor. In one embodiment, stage 1730 can be a linearly actuated stage that moves in a direction 1731, although it has been found that other types of stages and movement can be used in connection with the system described here. A set of mirrors 1752 is shown with respect to the second tube lens 1750 which may include one or more fold mirrors to adjust the light path from the second tube lens 1750 to the image sensor 1710.
[0182] Figures 39A and 39B are schematic illustrations of the 1700 optical duplication image system showing the transfer of the first 1740 tube lens and the second 1750 tube lens in front of the 1710 image sensor according to an embodiment of system described here. Figure 39A shows a 1741 light path for the first 1740 tube lens positioned in front of the 1710 image sensor at stage 1730. Figure 39B shows a 1751 light path for the second 1750 tube lens after being translated in front of the image sensor. image 1710 via stage 1730. As illustrated, the 1751 light path was increased using one or more mirrors from the 1752 mirror set. In both figures, it was found that the 1700 optical duplication image system can include other appropriate 1760 optical and structural components such as those discussed in detail elsewhere here.
[0183] Figure 40 shows an imaging system 2000 that has many components that are generally similar to the components shown in figures 1, 2, and 26. An access door 2002 can be opened to load cassettes on a carousel. The cassettes can carry laminated blades. After loading the cassettes, the access door 2002 can be closed. A 2004 controller can be used to operate the 2000 imaging system. After the samples have been imaged, door 2002 can be opened and the cassettes removed. Advantageously, slides can be imaged while the user loads and unloads the carousel to increase laboratory performance. This also avoids unnecessary waiting times and ensures that blades are always ready for processing.
[0184] A protective box 2010 from figure 40 is shown removed in figure 41. Figure 41 shows a 2011 caching device including a 2012 carousel with nine upper docking stations and nine lower docking stations. Each docking station is capable of receiving and securing a cassette or other type of blade holder device (for example, a blade holder, a cartridge, etc.). The 2012 carousel rotates to position cassettes in front of the 2000 system for convenient caster loading and unloading.
[0185] With respect to figures 41-43, blades can be moved to a temporary storage 2020 that features a 2022 return and a 2024 camera. A blade manipulator in the form of an XY Stage 2026 can be fed with blades through a manipulator 2028 blade optics. 2030 imaging optics can capture images of the sample on a slide loaded to stage 2026. Optics 2030 imaging optics can include without limitation one or more lenses, cameras, mirrors, filters, light sources, or the like. A 2034 gripping device or end effector carried by a 2036 arm can transport the blades between various components.
[0186] Figure 44 shows the clamping device 2034 that carries the 2050 slide without coming into contact with the peripheral area of a 2051 label and without coming into contact with the microscope slide edges adjacent to the 2051 label, thus preventing contact with exposed residual glue, if applicable, used to apply the 2051 tag. The 2050 blade can safely and efficiently transport blades without the problems caused by glue adhering to the 2034 grip device. As used here, the term “ slide ”includes a coverslip microscope slide as well as a coverslip microscope slide. The illustrated 2050 blade includes a coverslip. If the clamping device 2034 is part of a slide assembly device, the clamping device 2034 can load a microscope slide without a coverslip.
[0187] The clamping device 2034 includes a connector unit 2038 and a final effector 2040. The connector unit 2038 fluidly couples a top element 2044 to a pressure source (for example, a pump or other device capable of producing a vacuum). The top element 2044 is capable of maintaining a vacuum with the 2050 blade. A sufficient vacuum can be maintained to securely hold the 2050 blade and load the 2050 blade between workstations. To release the 2050 blade, the vacuum can be reduced or eliminated.
[0188] The connector unit 2038 includes a mounting body 2055 and a pair of mounting arms 2056a, 2056b attachable to the arm 2036 (see Figures 42 and 43). Fluidic components 2060 provide fluidic communication with the final effector 2040. Fluidic components include, without restriction, fluid line (eg hoses, tubes, conduits, etc.), valves, couplers (eg fluid line couplers), and other components for controlling or establishing a fluid flow. In figures 44 and 45, fluid component 2060 includes a feed line 2062 that is fluidly coupled to final effector 2040.
[0189] The connector unit 2038 can allow the final effector 2040 and / or microscope slide 2050 to strike an object and allow the final effector 2040 to deflect and return to its original position in order to inhibit, limit or substantially prevent damage to the final effector 2040 and / or blade 2050. A 2058 protective head can serve as a collision protection feature. The head 2058 is movably attached to the mounting body 2055. If the blade 2050 or end effector 2040, or both, come into contact with an object, the head 2058 can rotate as indicated by arrows 2059a, 2059b in figure 46, to avoid , limit or prevent damage to blade 2050 and / or end effector 2040. A thrust member 2061 can propel head 2058 to a seated position. When sufficient force is applied to the final effector 2040, the thrust force provided by the thrust member 2061 is overcome and the head 2058 moves with respect to the mounting body 2055. When the applied force is sufficiently low or substantially eliminated the limb from boost 2061 pushes the head 2058 back to the seated position. The 2061 drive member may include, without restriction, one or more springs (including spiral springs, coil springs, etc.), pneumatic drive members, or other components capable of providing a driving force.
[0190] With respect to figures 44 and 45, final effector 2040 includes a platform 2070, a fluid line 2072, and the top element 2044. Platform 2070 has an upper surface 2074 and a lower surface 2076 (Figure 46), and it can be made entirely or only partially of a rigid material (for example, a metal, a rigid plastic, etc.). The illustrated cantilever platform 2070 is mounted on the 2038 connector unit in a cantilever mode and can be made of sheet metal or metal such as stainless steel sheet.
[0191] With reference to figure 47, a 2089 sensor can include one or more circuits (for example, a flex circuit) with a relatively flat profile to analyze the microscope slide. To detect whether the blade is present, the 2089 sensor can be an optical sensor capable of analyzing reflective surfaces. As an example, the 2089 sensor can detect whether a slide is present by evaluating the reflectance, if any, of the slide label. The 2089 sensor can send an indicative signal if the blade is present. Based on the signal, controller 2004 can determine whether a blade is attached by the 2034 gripping device. Alternatively, sensor 2089 can be in the form of a barcode reader, scanner, contact sensor, proximity sensor , combinations of these or similar. Any number of sensors can be used to determine information associated with the 2050 microscope slide.
[0192] Spacers 2090a-d are coupled to the bottom surface 2076 and can physically come into contact with the 2050 microscope slide. As shown in figure 42, the 2090a-d spacers cooperate to secure the 2050 microscope slide in a substantially horizontal orientation. and substantially parallel to the platform 2070. A front spacer 2090c and a rear spacer 2090a are generally positioned along a longitudinal axis 2118 of platform 2070. Side displacement spacers 2090b, 2090d are positioned close to longitudinal sides 2091, 2093 of platform 2070. Any number of spacers that define different types of configurations can be used based on the desired interaction of the blade.
[0193] To avoid adhesion with excess residual glue, the 2090 spacers can be located in positions where the 2090a-d spacers do not come into contact with the glue. The 2090a spacer contacts the top of the 2051 tag and is spaced well away from the edges of the tag to ensure that it will not come into contact with the excess residual glue. Alternatively, spacer 2090a can be moved out of the label area in the direction of grip 2044 and still provide stability to the blade. Spacers 2090b, 2090d are positioned to contact an upper surface of the microscope slide at a location quite far from the edge of the 2051 label.
[0194] With reference again to Figures 44-46, fluid line 2072 is positioned above the upper surface 2074 and extends between the connector unit 2038 and top element 2044. To increase the stiffness of the final effector 2040, fluid line 2072 it may comprise a hypodermic tube made entirely or partly of steel (including stainless steel), nickel alloys or other suitable rigid material. In some embodiments, the outer diameter of the hypodermic tube may be less than approximately 0.3 inches. The gauge of the hypodermic tube can be located in a 7 gauge to 32 gauge range based on the Stubs needle gauge and be selected based on the desired vacuum to be extracted as well as the desired mechanical properties of the 2034 gripping device. Tubes of other sizes as well can be used. The position of the 2072 fluid line with respect to the 2070 platform can be selected to acquire a desired moment of inertia and overall profile. The fluid line 2072 can be moved away from the 2070 platform to increase the stiffness of the final effector 2040 by increasing the moment of inertia. To reduce the maximum height of the final effector 2040, the 2072 fluid line can be moved to be close to or in contact with the 2070 platform.
[0195] A fixture set 2077 features a fixture body 2078 for securing the fluid line 2072 and fixing elements 2080a, 2080b for opening and closing fixture 2077. To reduce the number of parts of the clamping device 2034, fluid line 2072 can be coupled (for example, adhered, glued, welded) directly to the 2070 platform.
[0196] With reference to figure 48, top element 2044 includes a connector 2100 and a suction head 2102. The connector 2100 is positioned on the upper side of platform 2070 and is coupled to the fluid line 2072. The suction head 2102 is positioned on the bottom side of the 2070 platform and is capable of maintaining a vacuum with the 2050 microscope slide blade. In some embodiments, the suction head 2102 can be made of a material (eg rubber, silicone, polymers, etc.). ) sufficiently deformable to maintain an airtight seal when a flap 2103 of the head 2102 covers an edge of the coverslip. This allows for a consistent and secure grip even if the suction head 2102 is out of alignment with the flap.
[0197] A positioner 2110 is keyed to the 2070 platform and can serve as an anti-rotation feature to keep the top element 2044 properly aligned with the 2070 platform even after a relatively long service life. The illustrated positioner 2110 is a protrusion positioned in a complementary cutout 2112. Additionally or alternatively, the top element 2044 can be attached to the platform 2070 using one or more glues or adhesives. In still other embodiments, mechanical fasteners (for example, screws, pins, or the like) can be used to mechanically couple the suction head 2102 to the 2070 platform.
[0198] The final effector 2040 may have a relatively flat profile in order to access relatively narrow spaces. Figure 48 shows a maximum height H of the final effector 2040 that can be in a range of approximately 0.15 inches to approximately 0.19 inches. This allows insertion of the final effector 2040 between microscope slides attached to a wide range of different types of slide holders, including slide doors with vertically spaced dividers. Such blade ports may be in the form of storage media, cartridges, portable cassettes and the like. The final effector 2040 can have different configurations, sizes, and dimensions and arrangement of selected components based on the desired application. The width W (see Figure 45) of the final effector 2040 can be equal to or less than the width of the blade 2050. In some embodiments, including an illustrated embodiment of figure 45, the final effector 2040 has a width W that is substantially less than the width of the 2050 blade. This allows side-by-side variation of the final effector 2040 with respect to the 2050 blade.
[0199] Figure 49 shows a microscope slide holder in the form of a 2400 cassette. Cassette 2400 includes a main body 2410 and vertically spaced dividers 2414. A lamella slide 2422 (shown in dashed line) is supported by a higher partition. 2414a. If the 2400 cassette is manually transported between workstations, a user can retain the 2400 cassette in a generally vertical orientation as shown. The blades can be conveniently retained in partitions 2414 even if cassette 2400 is tilted from the vertical orientation. The 2400 cassette can be portable for proper transport between workstations. Latches help prevent slides from sliding out of the 2400 cassette while the cassette is being transported (for example, when transporting the 2400 cassette between laboratory equipment) or due to mechanical vibrations induced, for example, by motor-driven equipment on a blade holder or close to the blade holder.
[0200] With respect to figures 49 and 50, the main body 2410 includes a front opening 2430, an upper wall 2444, a lower wall 2446, a rear wall 2448, and side walls 2452, 2454. The upper wall 1444 extends between the side walls 2452, 2454 and defines an access opening 2460. The bottom wall 2446 extends between the side walls 2452, 2454. A gripping device can pass through the access opening 2460 for loading and unloading blades. The bottom wall 2446 can rest on a support surface to keep the cassette 2400 in a generally vertical orientation. Handles 2447, 2449 can be attached to handle cassette 2400.
[0201] The rear wall 2448 includes ventilation openings 2470 through which the fluid (for example, air, mounting liquid, etc.) can pass. The illustrated openings 2470 are adjacent to the gaps between adjacent partitions 2414. If the blades are moist, air can pass through the openings 2470 for air drying or draining, or both. The lengths of the openings 2470 may be less than those widths of the blades to prevent the passage of blades through the back wall 2448. In addition or alternatively, ventilation openings may be in other locations such as along one or both side walls 2452, 2454 .
[0202] The main body 2410 can be made, in whole or in part, of plastic, polymers, metal or combinations of these and can have a construction in one piece or in multiple pieces. Representative non-restrictive plastics include, without limitation, acrylonitrile butadiene styrene (ABS), polyurethane, polyester, polypropylene, combinations thereof or the like. Loads can be used to increase performance. In some embodiments, the main body 2410 is made of ABS plastic that is filled with glass (for example, 30% filled with glass by volume or weight). The surfaces of partitions 1412 may have a relatively low coefficient of friction to allow convenient displacement of the blade placement. In molded embodiments, two molded halves can be welded, bonded or otherwise coupled together to form the 2400 cassette. Any number of molded portions can be assembled or coupled together. Alternatively, machined components can be assembled together to form the 2400 cassette. Mechanical fasteners, glue, solder or the like can be used.
[0203] A 2474 coupler in figure 50 is positioned at the rear of the 2400 cassette. If a workstation has a magnet (for example, a permanent magnet or an electromagnet), the 2474 coupler can be made in whole or in part from a ferromagnetic material or other material attached to magnets to help secure the 2400 cassette to the workstation. Alternatively, the 2474 coupler may include a magnet for coupling to a ferromagnetic component of the workstation.
[0204] Figure 50 also shows a pair of projecting switching elements 2482, 2484. When cassette 2400 is moved to a docking station, switching elements 2482, 2484 can be inserted into corresponding receivers (for example, grooves, openings, etc.) of the docking station. If the cassette 2400 is inserted inverted, the switching elements 2482, 2484 will not be received by receivers, thus avoiding fitting. The illustrated switching elements 2482, 2484 are generally U-shaped parallel members. The positions, configurations, and orientations of the switching elements can be selected based on the size of the cassette, desired cassette orientation, docking station configuration and similar. A spike 2485 of figure 49 can trigger a mechanical device (for example, a mechanical interlock) or a sensor that sends a signal indicating that the 2400 cassette is properly installed. In addition or alternatively, contact sensors, proximity sensors or other types of sensors can be located in various positions (for example, inside the 2012 carousel) to determine whether the 2400 cassette is properly loaded.
[0205] Figures 51 and 52 show twenty dividers vertically spaced 2414 apart for simulation of twenty slides. Each partition 2414 is positioned between side walls 2452, 2454 and oriented to support a microscope slide in a substantially horizontal orientation when the 2400 cassette is in the vertical position. Partitions 2414 may be spaced evenly or non-uniformly. In uniformly spaced embodiments, the average distances between adjacent partitions 2414 can be in the range of approximately 0.25 inches to approximately 0.38 inches. The average distance can be selected to acquire clearance for the grip head. The size, spacing and other dimensions of the partitions can be selected to facilitate transfer between blade retaining devices. As an example, the spacing of partitions 2414 in general can be similar to the spacing of partitions and grooves of a diving-type basket or buried so that blades can be conveniently inserted into the cassette from the basket. If blades are transferred from a slide holder to cassette 2400, partitions 2414 may have a spacing generally corresponding to the spacing of the holder partitions. In this way, the special arrangement of the 2414 partitions can be selected based on different processing parameters and criteria.
[0206] With respect to figure 51, openings are marked to conveniently locate a blade. The highest position opening 2490a is labeled with the letter "A," and the lowest position opening 2490t is labeled with the letter "T." Alternatively, openings can be labeled with numbers or other “clues”
[0207] Figures 51 and 52 show a coverslip blade for specimen support 2422 (illustrated in dashed line) on the partition 2414a. A pair of support members 2500a, 2500b supports a label end 2504 of blade 2422. Each support member 2500a, 2500b extends from partition 2414a. One unitary embodiments, support members 2500a, 2500b are integrally molded with the partition 2414a. In non-unitary embodiments, support members 2500a, 2500b are coupled to partition 2414a using adhesives, fasteners, male / female connectors or the like.
[0208] Support members 2500a, 2500b in general can be similar to each other and correspondingly, the description of one applies to the other, as long as there is no different indication. Figures 53 and 54 show the support member 2500a including an elongated body 2508a and a lock 2510a. The elongated body 2508a includes a mounting end 2516a coupled to the partition 2414, an opposite free end 2518a that carries the lock 2510a, and a central portion 2519a.
[0209] A shoulder 2522a of clasp 2510a protrudes upward from the elongated body 2508a and can limit the movement of the microscope slide 2422 with respect to the support member 2500a. The shoulder 2522a can obstruct the slide of the blade 2422. The height Hs of the shoulder 2522a can be less than the thickness TS defined by a mounting surface and a bottom surface of the blade 2422. In some embodiments, the height Hs is equal to or less than approximately 60%, 50%, or 40% of the TS thickness. In one embodiment, the height of the shoulder Hs is approximately half the thickness TS. Other boss heights are also possible if necessary or desired. The distance between lock 2510a and rear wall 2532 can generally be equal to or slightly longer than the longitudinal length of blade 2422. This can help to minimize the movement of blade 2422 during transport.
[0210] An anchor surface 2528a can generally be perpendicular to a support surface of the elongated body 2508a. In other embodiments, closure 2518 may be a burr, a protrusion (e.g., a partially spherical protrusion, ear, etc.), or other limiting element (e.g., inhibition or impediment) of movement of the blade 2504.
[0211] With respect to figure 55, partition 2414 has an H-shaped plate. The plate may have a generally flat top surface and in some embodiments, it is textured, polished or similar to increase the loading or unloading of blades for microscope. The illustrated partition 2414 is connected to and extends between side walls 2452 and 2454. In other embodiments, the partition 2414 is mounted in cantilever mode on one of the side walls.
[0212] The elongated body 2508a has a longitudinal axis 2540a that is substantially parallel to a longitudinal axis 2540b of an elongated body 2508b of a support member 2520b. The longitudinal axes 2540a, 2540b can be on a plane and can define an angle, if applicable, less than approximately 5 degrees. An illustrated embodiment has two support members, however additional support members can be used as well as any number of dividers.
[0213] Figures 56A and 56B show a microscope slide 2422 (shown in dashed line) being placed in the upper divider 2414. Guides 2560a, 2560b can help accommodate side-by-side variation of slide placement. As shown in figure 56A, blade 2422 is positioned to the left side of opening 2490a. An edge 2561a of blade 2422 can slide along a surface 2562a due to gravity to allow blade 2422 to fall in the desired location. The angled surfaces 2562a, 2562b are angled to allow the microscope slide to slide down and rest on the partition 2414a. In some embodiments, the angle of inclination of the surfaces 2562a, 2562b can in general be equal to one another.
[0214] With respect to Figure 40 again, to operate the 2000 imaging system, a user will be able to open the 2002 access door to insert cassettes into one of the eighteen docking stations (for example, grooves) of the 2012 carousel. The 2012 carousel it can then rotate for loading and unloading. Advantageously, 2012 carousel can have a relatively compact configuration to reduce the overall system footprint of the 2000 system.
[0215] Once a cassette is loaded into a docking station, a button 2602 (see Figure 41) can be turned counterclockwise (for example, rotated approximately 90 degrees counterclockwise) to lock the cassette on site. In addition, the rotation of button 2602 can produce rotation of a corresponding flag (see flag 2607) which is analyzed by a 2606 sensor. The position of the flag corresponds to the position of button 2602. Correspondingly, the 2606 sensor can determine whether the button is in an open position or a closed position.
[0216] The sensor 2606 can be an optical sensor capable of analyzing the position of three cassettes and three lower cassettes aligned.
[0217] In the illustrated embodiment, sensor 2606 determines the presence of cassettes 2400a-c based on the position of beacons connected to buttons 2602a-c. The lower 2400d-f cassettes are analyzed by another sensor. Advantageously, the combination of optical sensor / signaler allows a stationary column inside the center of the carousel to analyze whether the cassettes are present, thus avoiding the need for wires that cross the peripheral region of rotation and thereby eliminating the use of complicated cable holders . This can be useful to increase the reliability of the 2012 carousel. In some embodiments, the spike 2485 (see Figure 50) is used to engage a mechanical interlock that allows the 2602 buttons to rotate while the button on an empty docking station cannot be rotated.
[0218] With reference again to Figure 42, the blade manipulator 2028 moves the gripping device 2034 to transport blades between the various components of the 2000 imaging system. Figure 52 shows a flat end effector 2034 positioned to be inserted into opening 2490a, as indicated by an arrow 2616. Since the final effector 2034 is positioned above a microscope slide 2422, the final effector 2034 can be placed on a microscope slide 2422. A vacuum can be produced between the suction head 2102 and the microscope slide. The final effector 2034 can move the blade 2422 vertically close to the lock 2510a. The raised blade 2422 can be moved out of aperture 2490a. The final effector 2034 can load the 2422 slide to the desired location without coming in contact with glue (for example, glue on the microscope slide periphery or adjacent edges), the gripping device can move slides to any number of processing stations , including, without limitation, carousels, temporary storage, Stage XYs, blade manipulators, caching devices, media, imaging equipment, docking stations or the like. After a slide is imaged, the slide can be moved from the caching device 2020a to an empty cassette partition. Once a cassette is filled with imaged slides, the cassette can be rotated to the access region for convenient removal using the 2002 access door.
[0219] Various embodiments discussed here can be combined together in suitable combinations in connection with the system described here. In addition, in some cases, the order of steps in the flowcharts, flow diagrams and / or described flow processing can be modified when necessary. In addition, various aspects of the system described here can be implemented using software, hardware, a combination of software and hardware and / or other modules implemented by a computer or devices that have the characteristics described and perform the functions described. Software implementations of the system described herein may include executable code that is stored on a computer-readable storage medium and executed by one or more processors. The computer-readable storage medium may include a hard disk, ROM, RAM, flash memory, portable computer storage media such as CD-ROM, DVD-ROM, a flash drive and / or other drives with, for example, a universal serial bus interface (USB) and / or any other appropriate tangible storage medium or computer memory in which the executable code can be stored and executed by a processor. The system described here can be used in connection with any appropriate operating system.
[0220] Other embodiments of the invention will become apparent to the person skilled in the art from consideration of the specification or practice of the invention described herein. The specification and examples are here conceived as representative only, the true scope and spirit of the invention being indicated by the following claims.

权利要求:
Claims (7)
[0001]
1. MICROSCOPE SLIDE CARRYING CASSETTE (2400), comprising: a main body (2410) to surround and protect microscope slides (2422), the main body (2410) including a first side wall (2452) and a second side wall (2454); a plurality of partitions (2414) capable of supporting microscope slides (2422), the partitions (2414) being positioned between the first side wall (2452) and the second side wall (2454) and are vertically spaced apart from each other when a microscope slide cassette (2400) is in a vertical orientation; and a plurality of support members (2500a, 2500b) extending away from the respective partitions (2414), at least one of the support members (2500a) includes an elongated body (2508a) coupled to one of the partitions (2414) ), and a fastener (2510a) that extends upwards from the elongated body (2508a) and can limit the movement of a microscope slide (2422) positioned on the elongated body (2508a), characterized by the microscope slide carrier cassette ( 2400) further comprise a pair of guides (2560a, 2560b) close to one of the partitions (2414), wherein each of the guides (2560a, 2560b) includes an inclined surface (2562a, 2562b) distant from one of the partitions (2414) and provided for a microscope slide (2422) to slide due to gravity until the microscope slide (2422) rests on one of the dividers (2414), to accommodate side-by-side variation of slide placement in one of the dividers (2414 ).
[0002]
2. SLIDE CARRYING CASSETTE FOR MICROSCOPE, according to claim 1, characterized by the pairs of the support members (2500a, 2500b) being separated from each other and coupled to the respective partitions (2414), in which two of the elongated bodies (2508a) they are coupled to the respective partitions (2414) and have longitudinal axes (2540) that are parallel to each other.
[0003]
MICROSCOPE SLIDE CARRYING CASSETTE according to any one of claims 1 to 2, characterized in that the first side wall (2452) and the second side wall (2454) define an opening (2460) through which microscope slides ( 2422) can be inserted to place the microscope slides (2422) in the dividers (2414) and through which the microscope slides (2422) can be removed from the dividers (2414).
[0004]
4. MICROSCOPE SLIDE CARRYING CASSETTE according to any one of claims 1 to 3, characterized in that the elongated body (2508a) includes a mounting end (2516a) coupled to the partition (2414), an opposite free end (2518a) which it carries the lock (2510a), and a cantilever body (2519) that extends between the mounting end (2516a) and the free end (2518a).
[0005]
A MICROSCOPE SLIDE CARRYING CASSETTE according to any one of claims 1 to 4, characterized in that the closure (2510) is a protrusion adapted to provide a height equal to or less than half the thickness of a microscope slide (2422 ), when the microscope slide (2422) rests on the elongated body (2508a) and the partition (2414).
[0006]
6. MICROSCOPE SLIDE CARRYING CASSETTE according to any one of claims 1 to 5, characterized by at least one first side wall (2452), the second side wall (2454), and a rear wall (2448) including a or more ventilation openings (2470) adjacent to the gaps between adjacent partitions (2414).
[0007]
7. MICROSCOPE SLIDE CARRYING CASSETTE according to any one of claims 1 to 6, characterized in that it also comprises at least one switching element (2482, 2484) that protrudes from the main body (2410), and at least one element switching (2482, 2484) is receivable by a workstation when the cassette (2400) is in a vertical orientation.

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KR101869664B1|2018-06-20|

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SG10201600153YA|2016-02-26|

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JP2016006534A|2016-01-14|

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HK1219542A1|2017-04-07|

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EP2753968A1|2014-07-16|

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CA2844994C|2016-10-11|

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CN105204152B|2018-10-26|

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AU2012306572A1|2014-01-30|

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WO2013034430A1|2013-03-14|

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CN103765291A|2014-04-30|

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JP2014526712A|2014-10-06|

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AU2012306572B2|2015-04-23|

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法律状态:
2018-12-11| B06F| Objections, documents and/or translations needed after an examination request according [chapter 6.6 patent gazette]|

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2019-12-31| B06U| Preliminary requirement: requests with searches performed by other patent offices: procedure suspended [chapter 6.21 patent gazette]|

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2020-10-13| B09A| Decision: intention to grant [chapter 9.1 patent gazette]|

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2020-12-15| B16A| Patent or certificate of addition of invention granted|Free format text: PRAZO DE VALIDADE: 20 (VINTE) ANOS CONTADOS A PARTIR DE 21/08/2012, OBSERVADAS AS CONDICOES LEGAIS. |

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2021-01-12| B16C| Correction of notification of the grant|Free format text: REF. RPI 2606 DE 15/12/2020 QUANTO AO ENDERECO. |

优先权:

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申请号 | 申请日 | 专利标题

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US201161533114P| true| 2011-09-09|2011-09-09|

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US61/533,114|2011-09-09|

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PCT/EP2012/066266|WO2013034430A1|2011-09-09|2012-08-21|Imaging systems, cassettes, and methods of using the same|

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