专利摘要:
PCT No. PCT/GB87/00627 Sec. 371 Date Jul. 5, 1988 Sec. 102(e) Date Jul. 5, 1988 PCT Filed Sep. 7, 1987 PCT Pub. No. WO88/01736 PCT Pub. Date Mar. 10, 1988.Apparatus for optical analysis of particles such as biological cells utilizes twin beams passing through a common optical fibre to avoid problems of alignment. For scattering measurements, a narrower beam can be used to gate detection of a broader beam to avoid amiguities associated with off-axis particles. The twin beams can also be used for thermal lensing, fluorescence or other techniques. Different beam diameters can alternatively be achieved in a single beam by controlling the focal length of a beam focussing lens.
公开号:SU1743371A3
申请号:SU884356098
申请日:1988-05-04
公开日:1992-06-23
发明作者:Карр Роберт;Джон Кларк Дэвид;Эткинсон Тони
申请人:Паблик Хелт Лаборатори Сервис, Борд (Фирма);
IPC主号:
专利说明:

The invention relates to optical analysis, in particular to analysis using optical techniques of biological materials, such as protein or other macromolecules, cells, viruses, tissue fragments, etc.
Within a wide range of particle sizes, starting from particles smaller than a manometer and up to a millimeter, optical methods are most preferable for determining parameters such as particle size, concentration, particle shape and speed.
Methods of optical analysis are known that use the intensity of light scattered by individual particles as they pass through the illuminated scattering volume.
These methods assume an optical measurement volume, the configuration of which
small enough to allow individual particles to be examined in a sufficiently dilute suspension of particles. Using fine focusing of the laser beams, high intensity light can be concentrated into less than 1 µl measuring volumes.
It is known a device that operates using these principles. In this device information is obtained when the localized interference pattern of the particle under study crosses an interferogram is formed by two different sets of interfering rays of different wavelengths that are focused at two points spaced apart by a small distance in space. two lens dispersion systems. The interferograms are thus obtained in various sizes and
vj
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with XI
with
the internal interferogram is used as a trigger or a valve.
However, this device is cumbersome and difficult to adjust and is not suitable for use outside the laboratory.
The closest technical solution to the invention is a device whose principle of operation is based on the use of two laser beams with different wavelengths traveling concentrically, with one beam having a diameter smaller than the other. The inner beam passes in the central part of the outer beam. The particle passing through the inner narrow beam is at this moment located centrally relative to the outer beam and as a result, the scattered light signal of the inner beam is used as a trigger or a valve to collect the scattered light of the outer beam. In the absence of a light signal from the internal beam (this absence is an indication that the particle has not reached the center of the rays), the scattered light of the external beam is ignored. The signals from the two beams are spread using two detectors, adjusted so that each responds to only one wavelength. The known system includes two lasers emitting rays, which are then focused into the desired concentric configuration using a combination of lenses, mirrors, prisms and beam splitters. Each of these optical devices requires fine adjustment, often using micrometric regulators in three planes.
However, this device is cumbersome and requires careful handling. It is undesirable to use under conditions outside the laboratory and it cannot perform in-line analysis of remotely located samples.
The aim of the invention is to simplify the design and increase the operational characteristics by eliminating the need for beam alignment.
The drawing shows a block diagram of the proposed device.
A device for optically determining the size and number of suspended particles contains a laser source (not shown) of two probing beams differing in the length of a light-emitting wolf, a common optical fiber line for transmitting the first and second light beams, which coincide in direction but with different wavelength - light guide 1 , a common focusing lens 2 for focusing two rays at different focal points spaced longitudinally along an axis passing through the detection zone such that in the detection zone along erechnye dimensions of the two beams are in a predetermined ratio, as well as light detectors focused on
the detection zone and able to distinguish the light scattered from the first and second rays, respectively, deflecting system, including a focusing lens 3, a beam splitter 4 and two filters 5, installed to supply the scattered light of two rays to the measuring detector 6 and the valve detector 7. The detectors have a common power source 8 and feed signals to a multichannel analyzer 9 connected to the processor 10, which tracks the coincidence of signals in the measuring and valve channels, performs the appropriate analysis in the channel and measurements. Intermediate information is stored in memory unit 11.
Preferably, the transverse size of the second beam is significantly smaller than the transverse size of the first beam, and the detection of the scattered light of the second beam provides an indication that
that the particle fell into the detection zone and is located approximately in the center of the first beam.
In an advantageous embodiment, a common focusing lens 2 is attached to
free end of the fiber optic and includes a gradient index lens. In the case of using a microlens attached to the free end of the fiber optic fiber, it is possible to obtain
a light source that can form absolutely concentric measurement and valve beams and which is compact, durable, vibration resistant and adapted to
positioning at a remote location and possibly in adverse conditions.
A light beam with a wavelength H 1 (for example, 488 nm from an air-cooled argon laser) is introduced as a single
modes into the fiber optic fiber 1. A wave of a different wavelength Ar (for example, 632 nm from a helium-neon laser) is also introduced into the fiber 1 with the help of appropriate optical elements or at some intermediate point along its length with a fiber-optic connector. In another possible embodiment, the light may be introduced directly into the fiber optic light guide using a mounted on it
source in the form of a laser diode. Since the fiber optic light guide is one-mode only for a given wavelength, deviations from the Gaussian beam profile can occur at the second frequency.
Light with two wavelengths propagates through a fiber-optic fiber in the direction of its free end, where it travels the distance DI before falling into the gradient index microlens. Lens 2 can be mounted on the free end of an optical fiber using, for example, an epoxy compound. The installation may also include an element matching reflection coefficients. For example, the SELFOC 1.8 model lens proved to be suitable. The dispersion of the material from which the lens is made will, due to chromatographic aberration, cause the rays to focus on different distances F (Ai) n f (2z). The two rays 1 and 2 due to the intrinsic properties of the fiber optic line will be concentric. The ratio of the diameters of the beams, due to the different location of the focal points, will vary depending on the distance D2 from the microlens. By choosing the appropriate distance from the microlens, one can irradiate the scattering volume with two rays of different widths. The ratio of the width of the rays can thus be adjusted by choice. Similarly, the thickness of the rays themselves can be selected depending on the distance D2. By selecting the distance D2, it is possible to create positions when the internal beam is beam 1 or beam 2. It is preferable to choose a beam with a second wavelength as an internal beam, since a slight deviation from the Gauus profile is quite tolerable. The values of the distances F (R1) and P (R2) are changed simultaneously by adjusting the distance DL. It also allows you to change the ratio of the thickness of the rays for any given distance D2.
The deflection system includes a focusing lens 3, a beam splitter 4, and a pair of filters 5 installed to supply the scattered light of two rays to the measuring 6 and valve 7 detectors, respectively. The detectors have a common power supply 8 and feed signals to a multi-channel analyzer 9. The processor 10, which may be a commercially available microcomputer, monitors the coincidence of signals in the trigger channel and the measurement channel and performs the appropriate analysis of the signals in the measurement channel.
Since the measurement is made in cases where the scattered light is detected by the trigger beam, it is known that the particle is in the center of the measuring beam. Since only the inner beam is used as a trigger, it is not significant that the beam profile may deviate from the Gaussian one.
In addition, a fiber optic detector line can be applied to focus (using microlenses) each of the beams and to supply light to the separated photodetectors. A fiber optic light guide equipped with a gradient index lens, identical to that used to form a pair beam, is placed at an angle © and a distance equal to D2 from the concentric rays. The scattering volume examined by this fiber optic light guide,
will coincide with the shape of the beam at a given point and focus accurately with respect to each of the beams, and without much adjustment. This follows from the symmetry of the whole structure. Changing the angle © will change the scattering volume.
In the proposed construction, the end of the fiber optic light guide, provided with a lens, may be located in the wall of the pipeline, passing the sample samples, and, if necessary, at a certain distance from the lasers and signal processing equipment. If the detection system is made in the form of the same design, then the part of the whole device, located near the detection area, can be made extremely compact, strong and vibration-resistant, and the device itself is therefore acceptable for in-line processing. In the case of a specific application that requires analysis of particles in a number of different places, for example when analyzing a fermentation process, a number of fiber optic lines serving detectors or acting as sources
light, can be placed in optical contact with a single light source or signal processing device.
As well as the known devices, the proposed device can be used for
a number of different types of analysis. Particle sizes and distribution can be determined by analyzing the intensity of scattered light. By analyzing the signals associated with the intensity, depending on the size of the particles, it is possible to obtain accurate data on the distribution of particle size. The particle concentration can be determined by counting the number of registered light pulses per unit of time.
Particle velocity and velocity distribution can be investigated by determining the pulse width when a particle passes a given scattering volume.
In special applications, the scattered light can be examined for polarization and spectral characteristics. Particles can be specifically labeled with fluorescent molecules and dyes to isolate other particles from the population. Particle shape can be determined from polygonal light scattering (by analyzing the intensity) for a specific range of particle sizes.
权利要求:
Claims (3)
[1]
Claim 1. Device for optical determination of the size and number of suspended particles containing a laser source of two probe beams, differing in wavelength of light radiation, optically conjugated through elements of coaxial alignment and transport of beams with a counting volume, with which the first and second optically conjugate photodetectors for the selection of light scattered by particles at the first and second lengths respectively
waves whose outputs are connected to the first and second inputs of a multichannel analyzer, respectively, whose output is connected to a recorder, characterized in that, in order to simplify the design and improve performance by eliminating the need for beam alignment, as coaxial adjustment and transportation elements
a beam is used a fiber-optic light guide with a focusing lens, the plane of which is separated from the counting volume at a distance that allows in the region of the counting volume to distinguish the transverse dimensions
two bunches.
[2]
2. The device according to claim 1, characterized in that the focusing lens is attached to the free end of the optical fiber.
[3]
3. The device according to PP. 1 and 2, characterized in that the common focusing lens includes a gradient index lens.
/ Y / Y
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同族专利:
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EP0279840A1|1988-08-31|
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AT70127T|1991-12-15|
NO881949L|1988-07-04|
DK170033B1|1995-05-01|
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法律状态:
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