• 专利附录
  • 专利说明
  • 权利要求
  • 类似技术
  • 同族专利
  • 引用文献
  • 法律状态
  • 优先权
    • 专利摘要:
    A method of singularizing embryos is described, the method comprising providing a plurality of embryos (40) within a system (20) and sensing (34) at least one of the plurality of embryos in a liquid, and the method also comprising depositing (26 ) said at least one of the plurality of embryos on a surface (28).
    公开号:SE1051006A1
    申请号:SE1051006
    申请日:2010-09-29
    公开日:2011-03-31
    发明作者:Anthony P Swanda
    申请人:Weyerhaeuser Nr Co；
    IPC主号:
    专利说明:

    2 Washington, the disclosure of which is hereby incorporated by reference.
    At present, embryos are picked manually from a growing medium and physically placed on the plate for collection and insertion into an empty seed. Although such manual processes are effective, they are not without their limitations. As a non-limiting example, such manual operations are both labor intensive and time consuming and therefore expensive. As part of the process of producing large numbers of somatic embryos available for insertion into the produced seeds, it is desirable to minimize the manual labor element from the process.
    Summary The summary is provided to introduce a selection of concepts in a simplified form which are further described below in the detailed description. The summary is not intended to identify key features of what is stated in the claims, nor is it intended to be used as an aid in determining the scope of what is stated in the claims.
    A method for singularizing embryos is provided. The method comprises providing a plurality of embryos within a system and sensing at least one of the plurality of embryos in a liquid. The method also involves distributing at least one of the majority of embryos on a surface.
    Description of the Drawings The present aspect and many of the attendant advantages of this invention will be better understood by reference to the following detailed description taken in conjunction with the accompanying drawings in which: Fig. 1 is a diagrammatic view of an example of a system using a method; for singularizing embryos according to an embodiment of the present invention, Fig. 2A is a flow chart for a method for singularizing embryos according to an embodiment of the present invention, Fig. 2B is a continuation of the flow diagram in Fig. 2A, Fig. 3A is a flow chart of a method for singularizing embryos in accordance with another embodiment of the present disclosure, and Fig. 3B is a continuation of the flow chart of Fig. 3A.
    Detailed Description Fig. 1 schematically illustrates an automated system 20 for performing a method for singularizing embryos according to an embodiment of the present invention. The system 20 is suitably mounted in conjunction with a unit for mounting manufactured seeds (not shown) or is located at a distance from such a unit.
    The system 20 includes an embryo storage unit 22, a programmable logic controller (PLC) 24, a placement mechanism 26, and an embryo storage unit 28. The embryo storage unit 22 includes a singularization container 30, a lifting device 32, and a sensor 34. The singularization container 30 is suitably a container having a number of embryos 40 suspended in a liquid, such as a sterile nanopure (Nanopure) water. Preferably, the fluid is agitated to a sufficient degree to suspend all embryos 40. The singularization container 30 is mounted on the lifting device 32.
    The lifting device 32 includes a base plate 50 coupled to a well-known elevator 52, such as a screw elevator or a scissor elevator, to assist in maintaining a substantially constant pressure at the outlet of the singularization container 30. In the spirit of the present invention used in this context, the term "substantially" “Intended to include engineering-acceptable variations that result in almost constant fluid flow rates. Although an elevator 52 is used to assist in maintaining a substantially constant pressure, other devices known to maintain a substantial pressure are also acceptable. As a non-limiting example, a pump (not shown) may be placed in fluid communication with the singularization vessel 30 to maintain the substantially constant flow rate. Thus, such devices are acceptable equivalents and are within the scope of the present invention. Furthermore, while it is preferred to maintain a substantially constant pressure, a variable pressure is also within the scope of the present invention as described in more detail below.
    Embryos 40 are transported between the singularization container 30 and the locating mechanism 26 by fluid flowing through pipeline 60. The pipeline 60 extends between the singularization container 30 and the locating mechanism 26 and the sensor 34 is suitably located adjacent the pipeline 60 for sensing and / or detecting embryos 40 inside the pipeline 60, as described in more detail below.
    In the illustrative and exemplary embodiment, the flow rate of embryos 40 through the pipeline 60 is controlled by the elevator 52. Expressly, and as is well known, the flow rate within the pipeline 60 is proportional to the square root of the vertical distance between the outlet of the pipeline 60 at the placement mechanism 26 and the liquid level in the singularization container 30. As the liquid in the singularization container 30 decreases, the singularization container 30 is raised by the lifting mechanism 32. The elevator 52 raises the singularization container 30 at a fixed rate proportional to the flow rate inside the pipeline 60 to maintain a substantially constant flow.
    The pipeline 60 includes an inner diameter large enough to allow entry of individual embryos 40 to enter the pipeline 60 at any given time. Although multiple embryos 40 may be located longitudinally within the conduit 60, it is desirable that only a single embryo be able to enter the conduit 60 at any given time. It is also preferred that the conduit 60 be of a material, such as silicone, that is transparent or semi-transparent to allow detection of an embryo within the conduit 60 by the sensor 34.
    The sensor 34 is a well-known laser-based visual sensor used to detect when an embryo emerges from the singularization container 30. Such a sensor 34 is model no. LV-H300 / 100 series, manufactured and sold by Keyence Corporation in Osaka, Japan. The sensor is suitably mounted on the base plate 50 with the pipeline 60 operably arranged between components of the sensor 34. The sensor 34 is in turn in communication with the PLC 24.
    The system 20 may also include a second well known sensor (not shown) in communication with the singularization container 30. The second sensor is used to measure the hydrostatic pressure of the liquid in the singularization container 30. Such a sensor is a model no. FW-H07, manufactured and sold by Keyence Corporation in Osaka, Japan. Such a sensor uses ultrasonic waves to measure distances. Although an ultrasonic sensor is preferred, other types of sensors, including laser and radar based, are within the scope of the present invention. The second sensor is communication with PLC 24.
    The well-known PLC 24 suitably has a user interface for controlling the singularization process and the raising and lowering of the lifting device 32.
    One such PLC 24 is a DirectLOGlC 205 Modular Programmable Logic Controller (DL205 PLC), manufactured and sold by Koyo Electronics Industries Co., Ltd, Tokyo, Japan.
    PLC 24 is programmable to cooperate with the lifting device 32, the sensor 34, the second sensor and the positioning mechanism 26 during operation of system 20, as well as to allow the operator to adjust operating parameters. Operating parameters such as the number of embryos 40 placed on the embryo deposition unit 28, the distance between embryos 40 and the placement of embryos on the embryo deposition unit 28 can all be programmed as desired.
    PLC 24 can be programmed to control the distance and location of embryos 40 on embryo deposition unit 28 by tracking embryos as they flow through pipeline 60. In such an embodiment, PLC 24 includes a clock or timer and a register. One such register is an embryo location registry ("ELR"). ELR includes binary registers representing location along the length of pipeline 60. For example, ELR may divide pipeline 60 into fifty registers, which represent fifty sequential locations in pipeline 60. The first register location is suitably located closest to sensor 34 and the the last register is located at the end of the pipeline 60 where it connects to the locating mechanism 26. The ELR tracks and logs the path of the embryos as a function of time inside the pipeline 60, as described in detail below.
    The locating mechanism 26 includes a robot arm 80. Movement of the robotic arm 80 is controlled relative to the embryo deposition unit 28 to locate the outlet of the pipeline 60 over an open space on the embryo deposition unit 28. One such robotic arm is an ultra-movement robotic arm, model no. DA25- HT17-8 NO-B / 4, manufactured and sold by Ultramotion of Mattituck, New York.
    To provide the desired movement of the robot arm 80, the locating mechanism 26 also includes a well-known stepper motor (not shown), such as model no. PK266-E2.0A, manufactured and sold by Oriental Motor USA Corp. from Torrance, California.
    The robot arm 80 has two degrees of freedom to accommodate the exact placement of embryos 40 on the embryo deposition unit 28. In that regard, it is preferred that the robot arm 80 translate longitudinally along an axis indicated by the arrow 70. Furthermore, the robot arm moves the axis perpendicular to arrow 70, i.e. in and out from the side. The outlet of the pipeline 60 on the robot arm is suitably oriented at an angle relative to a vertical axis so that, when the fluid exits the pipeline 60, it is not perpendicular to the embryo deposition unit 28.
    It is also desirable that the robot arm 80 be controlled by PLC 24 in combination with ELR, sensor 34, and / or the other sensor. As a non-limiting example, if an embryo 40 is detected by the sensor 34, it sends a signal to the PLC 24 indicating the presence of the embryo. The signal is entered in the ELR as "true". If an embryo is not detected by the sensor 34, the register is "false". A "true" register is noted as "1" while a "false" register is noted as The number of registers in ELR is a function of the length of pipeline 60. For example, if pipeline 60 is 20 inches long and there are fifty registers, whereby each register represents 0.4 inches of the pipeline 60. Furthermore, in this example, the movement time of an embryo from the sensor 34 to the locating mechanism 26 is about one second. As a result, each register of the ELR represents a time of about 20 ms. The clock updates the register every 20 ms, so that the register is shifted forward and each register is updated with “1” or Furthermore, the robot arm's 80 speed is also updated every 20 ms and is programmed to match the distance between the embryos, as described by the operator to control the distance between the embryos deposited on the embryo deposition unit 28.
    The embryo deposition unit 28 includes a singularization frame 82 and a drainage container 84. The singularization frame 82 suitably includes a support material that allows fluid to pass through while embryos are being retained. The support material also preferably provides a color contrast between the support material and the embryo so that there is a contrast between the embryo and the support material. One such support material for use with the system 20 is Nitex® nylon, model no. 03-125 / 45. The drainage container 84 suitably supports a vacuum (not shown) for fluid removal and for holding the embryo in a fixed position.
    Operating aspects of the system 20 constructed in accordance with an embodiment of the present invention are best understood with reference to Figs. 2A-2B. The beginning of the operating sequence is represented by the start block 100 by initializing the system 20 to zero in the ELR, indicated by block 102. Liquid flow through the system 20 is also initiated and the lifting device 32 lifts the singularization container 30 at a rate to maintain a substantially constant fluid pressure through the system 20. This is illustrated by block 104.
    The timer is activated, indicated by block 110, and the sensor 34 determines if the embryo 40 is detected in the pipeline 60 and indicated by the decision block 106.
    If an embryo 40 is detected by the sensor 34, “1” is placed on the first register location in the ELR, indicated by block 108. Thereafter, the timer is evaluated to determine if a predetermined time, such as 20 ms, has exceeded the time or not, which is indicated by decision block 112. If an embryo is not detected by sensor 34, the PLC advances forward to block 112 and evaluates whether the timer has exceeded the time.
    If the timer has not exceeded the time, the ELR returns to block 106 to evaluate whether an embryo has been detected. If the timer has exceeded the time, the register switches with a forward position, indicated by block 114. As indicated by block 116, the timer is also reset.
    As indicated by block 118, the PLC evaluates whether there is a “1” in the last register, which indicates the presence of an embryo 40 at the end of the pipeline 60. If there is a “0” in the last register, which indicates that if there is no embryo in the most recent register, the PLC determines if each register of the ELR is indicated by block 120. If each register is empty, the robot arm 80 is turned off, as indicated by block 122, and the PLC returns to block 110 to activate the increase timer and evaluate whether an embryo is detected by the sensor 34, as indicated by block 106. 10 15 20 25 30 8 Referring back to block 118, if the last register in ELR contains then PLC evaluates if another register in ELR contains thus indicating the presence of another embryo in the pipeline 60. This is indicated by block 124. As represented by 126, if no other register in the ELR contains it, the speed of the robot arm 80 is set to a minimum. This can be accomplished by including a look-up table containing predetermined robot arm velocities as a function of the number of embryos in the pipeline 60. Such a look-up table is well known to those skilled in the art.
    If it is “1” in one or more other registers of the ELR, the PLC sets the robot arm speed based on the last and penultimate register positions in the ELR by referring to the look-up table, as above. This is indicated by block 128. Then, as indicated by block 130, the output speed is sent to the robot arm 80.
    Before depositing embryos on the singularization frame 82, position "X" of the robot arm 80 is evaluated relative to the width of the singularization frame 82.
    Specifically, as indicated by block 132, position "X" of the robot arm 80 is evaluated to determine if it has reached the maximum width of the singularization frame 82. If so, the robot arm 80 advances one position forward in the longitudinal direction, or " The Y ”direction of the singularization frame 82 and the direction of the robot arm 80 are reversed, as indicated by block 134.
    After position "X" of the robot arm 80 is reserved, the PLZ zeros the position indicated by block 136. Thereafter, the embryo is deposited on the singularization frame 82, as indicated by block 138. Returning to block 132, if position "X" is not reached, blocks 134 and 136 are skipped and the embryo is deposited on the singularization frame 82, according to block 138.
    It is preferred that PLC 24 be programmed to control the robot arm 80 so that it deposits embryos at a predefined position on the singularization frame 82. As a non-limiting example, PLC 24 may be programmed so that the robot arm 80 deposits embryos on the singularization frame 82 on its sides. In such a position, both the heart leaf and the root canal terminals have contact with the support material of the singularization frame 82. As another non-limiting example, the robot arm 82 may deposit embryos on the support material so that subsequent embryos are spaced from previous embryos. such predetermined positions, as well as those corresponding thereto, are within the scope of the invention.
    After embryos are deposited on the singularization frame 82, indicated by block 140, the ELR determines whether a preferred number of embryos deposited on the singularization frame 82 have been achieved. If "no", the ELR returns to block 110 and the evaluation is repeated. If the maximum number of embryos has been deposited on the singularization frame 82, the process is complete, as indicated by block 142.
    Operation of an alternative method of singularizing embryos can best be understood with reference to Fig. 3A and Fig. 3B. It should be noted that components for this alternative embodiment which are the same as those described with respect to the first embodiment in Figs. 3A and 3B have the same reference numerals.
    The beginning of the operating sequence is represented by start block 100 by initiating system 20 with "0" in ELR, indicated by block 102. At the same time, fluid flow is initiated through system 20 and indicated by block 204. An increase timer is started, indicated by block 206, and the singularization rate, or the data point, is calculated, as indicated in block 208.
    The singularization rate is compared with the setpoint to determine whether or not the embryonicularization rate is equal to the setpoint, as indicated by decision block 210. The singularization rate is defined as the number of embryos detected per unit time. To calculate it, the number of detected embryos in a moving time window is divided by the size (in time) of the time window, e.g. 50 detections in the last 60 seconds. The window "moves" forward in time, as the last window is always used. If the embryosularization rate is not equal to the setpoint, the hydrostatic setpoint is adjusted. If the rate of embryosularization must be reduced, the hydrostatic setpoint is reduced. This is indicated by block 212, when the hydrostatic pressure of the liquid inside the singularization vessel 30 is measured by the second sensor. Such a sensor is described above. This is indicated by block 214.
    Still referring to Fig. 3A, a comparison of the hydrostatic pressure relative to the setpoint is made to determine if the hydrostatic pressure is equal to the setpoint or not, as indicated by block 216. If the hydrostatic the pressure is not at the set point, the lifting speed of the singularization container 30 is changed by the lifting mechanism 32, as indicated by block 218.
    In summary, the singularization velocity controller adjusts the hydrostatic setpoint (i.e., target fluid / embryo flow rate) and the hydrostatic pressure controller adjusts the rate of increase of the singularization vessel in an attempt to drive the hydrostatic pressure toward its target (setpoint). Following adjustment of the hydrostatic pressure, the length (ie the number of registers) of the ELR is calculated as indicated by block 220. The length of the ELR is calculated based on the distance between the sensor 34 and the outlet of the pipeline 60 and the flow rate of liquid (ie hydrostatic print). As the flow rate (pressure) increases, the velocity of fluid / embryos in the pipeline 60 increases, which in turn reduces the time between detection and placement on the s-frame 82. The number of registers required is this time divided by the time of the timer 2 in blocks 220. After block 220, a second boost timer is activated, as shown in block 221.
    The sensor 34 determines whether an embryo 40 has been detected in the pipeline 60 or not and is indicated by decision block 106.0. If an embryo 40 is detected by the sensor 34, "1" is placed in the first register location in the ELR, indicated by block 108. The second increase timer is then evaluated to determine if a predetermined time period, such as 20 milliseconds, has elapsed, as indicated by decision block 222. If an embryo is not detected by sensor 34, the PLC advances to block 222 to determine if the second gain timer has exceeded the time.
    If the second augmentation timer has not exceeded its time, the ELR returns to block 106 to evaluate if an embryo has been detected. If the second timer has exceeded its time, the ELD register shifts one position forward and puts “1” in the next register position, indicated by block 114. The second timer is also reset, as indicated by block 116.
    As indicated by decision block 118, the PLC evaluates whether it is “1” in the most recent or “trigger” ELR register, indicating the presence of an embryo 40 at each end of the pipeline 60. If it is “0” in the most recent register, indicating that there is no embryo in the last or 'trigger' register, the PLC determines if the first increase timer has exceeded its time, indicated by decision block 10 15 20 25 30 11 224. If the first increase timer has not exceeded its time, then the PLC advances back to activate the second boost timer, indicated by block 221.
    However, if the first increment timer has exceeded its time, the PLC returns to activate Timer1, indicated by block 206. Returning to decision block 118, if the most recent or trigger register in the ELR contains, the PLC deposits an embryo on the singularization frame 82, according to block 138.
    After depositing the embryo on the singularization frame 82, the position of the robotic arm relative to the width of the singularization frame 82 is evaluated.
    The "X" position of the robot arm is evaluated separately, as indicated by block 132, to determine if it has reached the maximum width of the singularization frame 82. If it has reached the maximum width of the singularization frame 82, the robot arm is advanced one position forward in the longitudinal direction, or the "Y" direction of the singularization frame 82, and the direction of the robot arm 80 in the "X" direction is reversed, as indicated by block 134. After the "X" position of the robot arm is reversed, the PLC zeroes out. The "X" position, indicated by block 136.
    If the “X” position is not reached in block 132, the robot arm is moved one position along the “X” axis, as indicated by block 226. By doing so, the robot arm 80 is moved to the next open position on the singularization frame 82. Thus, removal of at least one of the majority of embryos is synchronized with the data point, such as the hydrostatic pressure and the fate rate.
    Thereafter, the PLC determines if a desired number of embryos deposited on the singularization frame 82 has been reached, as indicated by block 140. If the desired number of counted embryos has been reached, the program returns to block 204 and the process is repeated. If the maximum number of embryos has been deposited on the singularization frame, the process is complete, as indicated by block 142. Although illustrative embodiments have been illustrated and described, it is to be understood that various changes may be made without departing from the spirit and scope of the invention. As a non-limiting example, the sensor 34 may be located at any point along the pipeline 60. In an alternative embodiment, the sensor 34 may be located adjacent the robot arm 80. In such an alternative embodiment, the PLC 24 may be programmed to activate the robot arm 80 to deposit the sensed embryo as soon as it receives an input signal from the sensor 34. Placing the sensor 34 adjacent to the robot arm 80 operates in a system 20 that has either constant or non-constant fluid flow. The method according to the present invention can also be implemented on a number of different systems and therefore the described system for implementing the method is provided for illustrative purposes only and is not intended to be limiting.
    权利要求:
    Claims (20)
    [1]
    A method of singularizing embryos, comprising: (a) providing a plurality of embryos, (b) detecting at least one of the plurality of embryos in a fluid, (c) tracking said at least one of the plurality of embryos as it is transported by the fluid, and (d) depositing said at least one of the plurality of embryos on a surface as a function of fluid flow rate.
    [2]
    The method of claim 1, further comprising maintaining a substantially constant flow rate while the at least one of the plurality of embryos is tracked as it is transported by the fluid.
    [3]
    The method of claim 1, wherein the tracking of said at least one of the plurality of embryos when transported by the fluid comprises a sensor.
    [4]
    The method of claim 1, wherein the sensor is in communication with a central unit for controlling the deposition of said at least one of the plurality of embryos on a surface.
    [5]
    The method of claim 1, wherein depositing said at least one of the plurality of embryos on a surface as a function of fluid flow rate comprises placing said at least one of the plurality of embryos at a predetermined position on the surface.
    [6]
    A method of singularizing embryos, comprising: (a) providing a plurality of embryos in a fluid, (b) sensing at least one of the plurality of embryos with a sensor, (c) maintaining a substantially constant flow rate of embryos past the sensor, and (d) depositing said at least one of the plurality of embryos on a surface. 10 15 20 25 30 14
    [7]
    The method of claim 6, wherein said at least one of the plurality of embryos is deposited at a predetermined position on the surface.
    [8]
    The method of claim 7, wherein the predetermined position comprises placing said at least one of the plurality of embryos on its side.
    [9]
    The method of claim 7, wherein the predetermined position comprises separating said at least one of the plurality of embryos from a second embryo located on the surface.
    [10]
    A method according to claim 6, wherein the sensor is in communication with a central unit for controlling the deposition of said at least one of the emb numbered embryos on a surface.
    [11]
    The method of claim 10, wherein the depositing of at least one of the plurality of embryos on a surface comprises a robotic arm in communication with the central unit.
    [12]
    A method according to claim 11, wherein the central unit is programmable to control movement of the robot arm.
    [13]
    The method of claim 12, wherein the movement of the robotic arm is programmed as a function of the substantially constant fluid flow rate.
    [14]
    A method of singularizing embryos, comprising: (a) providing a plurality of embryos in a storage container, (b) occupying a data point associated with the storage container, (c) determining a flow rate of embryos leaving the storage container, and (d) synchronize the removal of the at least one of the tal ertal embryos as a function of the data point and the flow rate. 10 15 20 15
    [15]
    The method of claim 14, wherein recording the data point comprises monitoring the storage container to determine a hydrostatic pressure.
    [16]
    The method of claim 14, wherein the synchronizing the removal of said at least one of the plurality of embryos comprises raising the storage container to increase the flow rate of embryos leaving the storage container.
    [17]
    The method of claim 14, wherein the synchronizing the removal of said at least one of the plurality of embryos comprises lowering the storage container to reduce the flow rate of embryos leaving the storage container.
    [18]
    The method of claim 14, wherein the synchronizing the removal of said at least one of the plurality of embryos comprises comparing the data point with a predetermined set of points.
    [19]
    The method of claim 18, wherein the data point is a hydrostatic pressure.
    [20]
    The method of claim 14, further comprising depositing said at least one of the plurality of embryos on a surface.
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    法律状态:
    2015-03-31| NAV| Patent application has lapsed|
    2015-04-28| CORR| Corrections|Free format text: PA GRUND AV ETT MISSTAG HAR ANSOEKAN 1051006-3 PUBLICERATS SOM "ALLMAENT TILLGAENG LIG PATENTANSOEKAN SOM SLUTBEHANDLATS UTAN ATT LEDA TILL PATENT". ANSOEKAN AER OFF ENTLIG MEN AENNU INTE SLUTLIGT AVGJORD. |
    2018-06-05| NAV| Patent application has lapsed|
    优先权:
    申请号 | 申请日 | 专利标题
    US24704709P| true| 2009-09-30|2009-09-30|
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