CN109060749B - Optical device for cell counting and detection - Google Patents
Optical device for cell counting and detection Download PDFInfo
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- CN109060749B CN109060749B CN201810961915.7A CN201810961915A CN109060749B CN 109060749 B CN109060749 B CN 109060749B CN 201810961915 A CN201810961915 A CN 201810961915A CN 109060749 B CN109060749 B CN 109060749B
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- 238000007493 shaping process Methods 0.000 claims abstract description 36
- 230000005540 biological transmission Effects 0.000 claims abstract description 21
- 230000005499 meniscus Effects 0.000 claims description 19
- 239000013307 optical fiber Substances 0.000 claims description 15
- 239000012530 fluid Substances 0.000 claims description 4
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- 238000005516 engineering process Methods 0.000 description 1
- 230000007274 generation of a signal involved in cell-cell signaling Effects 0.000 description 1
- 238000003384 imaging method Methods 0.000 description 1
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- G—PHYSICS
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- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/62—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
- G01N21/63—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
- G01N21/64—Fluorescence; Phosphorescence
- G01N21/645—Specially adapted constructive features of fluorimeters
- G01N21/6456—Spatial resolved fluorescence measurements; Imaging
- G01N21/6458—Fluorescence microscopy
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/62—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
- G01N21/63—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
- G01N21/64—Fluorescence; Phosphorescence
- G01N21/645—Specially adapted constructive features of fluorimeters
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Abstract
The invention discloses an optical device for counting and detecting cells, wherein a sheath flow device is positioned on the light path junction of 3 units of a light source shaping unit, a fluorescent signal receiving unit and 0-degree and 10-degree signal receiving units; the light source shaping unit comprises a light source shaping unit, a sheath flow device, a signal receiving unit, a light guide unit, a fluorescent signal receiving unit, a light source shaping unit, a fluorescent signal receiving unit and a fluorescent signal receiving unit, wherein the light emitting end of the light source shaping unit is connected with the incident end of the sheath flow device, one of the light emitting ends of the sheath flow device is connected with the incident end of the signal receiving unit at 0 degrees and 10 degrees, and the other light emitting end of the sheath flow device is connected with the incident end of the light guide unit. By designing the optical device of the cell analysis instrument, the invention improves the utilization efficiency of excitation light and fluorescence transmission efficiency, shortens the transmission distance of fluorescence, reduces the loss of fluorescence signals, and improves the utilization rate of light, thereby improving the detection accuracy.
Description
Technical Field
The invention relates to the technical field of cell analysis, in particular to an optical device for cell counting and detection.
Background
Currently, the instruments for cell counting and detection include a flow cytometer, a hemocytometer, a leucocyte classification detector and the like, and the instruments are all the cell analysis instruments for performing multi-parameter rapid quantitative analysis such as cell volume, cell internal complexity and the like based on a light scattering technology so as to analyze and sort specific cell populations. The working principle of these cell analysis instruments is: the laser light source performs shaping focusing through a plurality of lenses, compresses the light spot size to a certain size range, irradiates the laser beam to the capillary holes in the sheath flow device through the reflecting mirror, generates scattered light signals when the laser beam irradiates cells in the capillary holes through the sheath flow device, then collects, focuses and collimates the light signals through the lenses, propagates the collected light signals to the photomultiplier or the photodiode to receive and convert the light signals, and sends the electric signals to the upper computer to perform a series of signal processing to realize the characteristic analysis of the cells. Because the upper computer is used for analyzing the cells based on the cell scattered light signals, the light path system of the cell analysis instrument determines the performance of the cell analysis instrument, and the more reasonable the light path system is designed, the smaller the cell scattered light signal loss, the higher the signal-to-noise ratio of the cell scattered light signals, and the more accurate the detection result of the cell analysis instrument.
Disclosure of Invention
The invention aims to provide an optical device for cell counting and detection, which can reduce the loss of scattered light signals of cells, improve the signal-to-noise ratio of the scattered light signals and realize the accurate detection of cell structures and internal characteristics.
In order to solve the problems, the invention is realized by the following technical scheme:
an optical device for counting and detecting cells mainly comprises a base, a light source shaping unit, a sheath flow device, 0-degree and 10-degree signal receiving units, a light guide unit and a fluorescent signal receiving unit, wherein the light source shaping unit, the sheath flow device, the 0-degree and 10-degree signal receiving units, the light guide unit and the fluorescent signal receiving unit are arranged on the base; the sheath flow device is positioned on the light path junction of the light source shaping unit, the fluorescent signal receiving unit and the 3 units of the 0-degree and 10-degree signal receiving units; the light source shaping unit comprises a light source shaping unit, a sheath flow device, a signal receiving unit, a light guide unit, a fluorescent signal receiving unit, a light source shaping unit, a fluorescent signal receiving unit and a fluorescent signal receiving unit, wherein the light emitting end of the light source shaping unit is connected with the incident end of the sheath flow device, one of the light emitting ends of the sheath flow device is connected with the incident end of the signal receiving unit at 0 degrees and 10 degrees, and the other light emitting end of the sheath flow device is connected with the incident end of the light guide unit.
In the scheme, the fluorescent signal receiving unit consists of a signal receiving shell, and 5 fluorescent dichroic mirrors and 6 photoelectric converters which are arranged in the signal receiving shell; the incident end of the first fluorescent dichroic mirror forms the incident end of the fluorescent signal receiving unit; the reflection output end of the first fluorescent dichroic mirror is connected with the first photoelectric converter; the transmission output end of the first fluorescent dichroic mirror is connected with the incident end of the second fluorescent dichroic mirror; the reflection output end of the second fluorescent dichroic mirror is connected with the incidence end of the fifth fluorescent dichroic mirror, and the transmission output end of the second fluorescent dichroic mirror is connected with the incidence end of the third fluorescent dichroic mirror; the reflection output end of the fifth fluorescent dichroic mirror is connected with the sixth photoelectric converter; the transmission output end of the fifth fluorescent dichroic mirror is connected with a fifth photoelectric converter; the incident output end of the third fluorescent dichroic mirror is connected with the incident end of the fourth fluorescent dichroic mirror; the transmission output end of the third fluorescent dichroic mirror is connected with the second photoelectric converter; the reflection output end of the fourth fluorescent dichroic mirror is connected with the third photoelectric converter; the transmission output end of the fourth fluorescent dichroic mirror is connected with a fourth photoelectric converter.
As an improvement, the output end of the fluorescent dichroic mirror is connected with the photoelectric converter through a bandpass filter.
As an improvement, the fluorescent dichroic mirror is removably mounted in the signal receiving housing.
In the scheme, the light guide unit consists of a light guide base, a front light guide meniscus lens, a rear light guide meniscus lens, a double-cemented lens, a microscope objective lens cone, a slit baffle, a plano-concave microscope eyepiece and a plano-convex microscope eyepiece; a light guide through hole is formed in the light guide base; an entrance of the light guide through hole forms an incident end of the light guide unit and faces the sheath flow device; the outlet of the light guide through hole forms an emergent end of the light guide unit and faces the fluorescent signal receiving unit; the front light guide meniscus lens, the rear light guide meniscus lens, the double-cemented lens and the microscope objective lens are sequentially arranged at the entrance of the light guide through hole from front to back, the front light guide meniscus lens is arranged at one side close to the entrance of the light guide through hole, and the microscope objective lens is arranged at one side close to the exit of the light guide through hole; the slit separation blade, the plano-concave microscopic ocular and the plano-convex microscopic ocular are sequentially arranged at the outlet of the light guide through hole from front to back, the slit separation blade is arranged at one side close to the inlet of the light guide through hole, and the plano-convex microscopic ocular is arranged at one side close to the outlet of the light guide through hole.
In the above scheme, the light guide unit is an optical fiber bundle, one end of the optical fiber bundle is coupled with the sheath current device, and the other end of the optical fiber bundle is coupled with the incident end of the fluorescent signal receiving unit.
As an improvement, a plano-convex lens is additionally arranged between the optical fiber bundle and the sheath flow device.
In the scheme, the 0-degree and 10-degree signal receiving unit consists of a first receiving base, a diaphragm, a forward collecting lens, a perforated lens, a 0-degree signal receiving circuit board and a 10-degree signal receiving circuit board; the first receiving base is internally provided with a light inlet through hole, a 0-degree light outlet through hole and a 10-degree light outlet through hole which are communicated; the 0-degree light-emitting through hole is coincided with the central axis of the light-entering through hole, the 10-degree light-emitting through hole is perpendicular to the central axis of the light-entering through hole, and the central axes of the light-entering through hole, the 0-degree light-emitting through hole and the 10-degree light-emitting through hole are intersected at the intersection of the 3 through holes; the perforated mirror is arranged at the junction, and the outlet of the light inlet through hole is connected with the inlet of the 0-degree light outlet through hole and the inlet of the 10-degree light outlet through hole through the perforated mirror; the entrance of the light-in through hole forms the incident end of the 0-degree and 10-degree signal receiving unit, and faces the sheath flow device, and the forward condensing lens is arranged at the entrance of the light-in through hole; the 0 degree signal receiving circuit board is arranged at the outlet of the 0 degree light emitting through hole, and the 10 degree signal receiving circuit board is arranged at the outlet of the 10 degree light emitting through hole.
As an improvement, a diaphragm is arranged at the front side of the front condenser.
In the scheme, the light source shaping unit mainly comprises 2 lasers, 4 cylindrical mirrors, 1 reflecting mirror, 1 light source dichroic mirror and 1 light source focusing lens; the 2 lasers have different wavelengths, wherein one laser enters the reflecting mirror after passing through 2 mutually orthogonal cylindrical mirrors and enters one incident end of the light source dichroic mirror after being reflected by the reflecting mirror; the other laser directly enters the other incident end of the light source dichroic mirror after passing through 2 mutually orthogonal cylindrical mirrors; the emergent end of the light source dichroic mirror is connected with the incident end of the light source focusing lens, and the emergent end of the light source focusing lens forms the emergent end of the light source shaping unit.
As an improvement, the sheath flow device is arranged on the base through an azimuth adjusting plate which can realize the adjustment of X-axis, Y-axis and Z-axis directions.
Compared with the prior art, the invention has the following characteristics:
1. by designing the optical device of the cell analysis instrument, the utilization efficiency and fluorescence transmission efficiency of excitation light are improved, the transmission distance of fluorescence is shortened, the loss of fluorescence signals is reduced, the utilization rate of light is improved, and the detection accuracy is improved.
2. The dual excitation light is adopted to detect the cells, so that various characteristics of one cell can be analyzed, and the detection efficiency is improved;
3. the fluorescent dichroic mirror of the fluorescent signal receiving unit adopts a pluggable design, so that a plurality of fluorescent channels can be replaced freely, channel configuration is changed conveniently, and the fluorescent dichroic mirror is flexibly updated, and the use cost and the maintenance cost are saved for users greatly;
4. the light guide unit adopts the light transmission type optical fiber to collect scattered fluorescent signals, so that the light path structure can be reduced to a certain extent, the size of the instrument is miniaturized, the cost is lower, and the low loss of the optical signals is realized by utilizing the total reflection characteristic of the optical fiber.
Drawings
FIG. 1 is a schematic perspective view of an optical device for cell counting and detection;
FIG. 2 is a top view of an optical device for cell counting and detection;
FIG. 3 is a schematic diagram of the optical path of an optical device for cell counting and detection;
fig. 4 is a schematic perspective view of a light source shaping unit;
FIG. 5 is a perspective cross-sectional view of the 0 degree and 10 degree signal receiving units;
fig. 6 is a perspective cross-sectional view of a light guiding unit (first embodiment);
fig. 7 is a cross-sectional view of the fluorescent signal receiving unit (photoelectric converter removed);
FIG. 8 is a schematic diagram of the optical path of a fluorescent signal receiving unit;
fig. 9 is a schematic diagram of another installation of the light guide unit (second embodiment);
reference numerals in the drawings: 1. a base; 2. the device comprises a light source shaping unit, a 2-1 laser, 2-2 cylindrical mirrors, 2-3 reflecting mirrors, 2-4 light source dichroic mirrors, 2-5 light source focusing lenses; 3. a sheath flow device; 4. the device comprises 0-degree and 10-degree signal receiving units, 4-1 receiving bases, 4-1-1 receiving base, light entering through holes, 4-1-2 receiving base, 0-degree light emitting through holes, 4-1-3 receiving base, 10-degree light emitting through holes, 4-2 receiving base, diaphragms, 4-3 receiving base, a forward collecting lens, 4-4 receiving base, a perforating lens, 4-5 receiving base, 0-degree signal receiving circuit boards and 4-6 receiving base and 10-degree signal receiving base boards; 5. the light guide unit comprises 5-1 parts of light guide units, a light guide base, 5-1 parts of light guide through holes, 5-2 parts of front light guide meniscus lenses, 5-3 parts of rear light guide meniscus lenses, 5-4 parts of double-cemented lenses, 5-5 parts of microscope objective lens barrels, 5-6 parts of microscope objective adjusting parts, 5-7 parts of slit baffle plates, 5-8 parts of plano-concave microscope eyepieces, 5-9 parts of plano-convex microscope eyepieces, 5-10 parts of optical fiber bundles; 6. the fluorescent signal receiving unit comprises a fluorescent signal receiving unit 6-1, a signal receiving shell, a fluorescent dichroic mirror 6-2, a fluorescent dichroic mirror 6-3, a band-pass filter, a photoelectric converter 6-4.
Detailed Description
The invention will be further described in detail below with reference to specific examples and with reference to the accompanying drawings, in order to make the objects, technical solutions and advantages of the invention more apparent. In the examples, directional terms such as "upper", "lower", "middle", "left", "right", "front", "rear", and the like are merely directions with reference to the drawings. Accordingly, the directions of use are merely illustrative and not intended to limit the scope of the invention.
Embodiment one:
referring to fig. 1-3, an optical device for counting and detecting cells mainly comprises a base 1, a light source shaping unit 2, a sheath flow device 3, 0-degree and 10-degree signal receiving units 4, a light guiding unit 5 and a fluorescent signal receiving unit 6, wherein the light source shaping unit 2, the sheath flow device 3, the 0-degree and 10-degree signal receiving unit 4, the light guiding unit 5 and the fluorescent signal receiving unit 6 are arranged on the base 1. The sheath fluid device 3 is positioned at the light path junction point of the light source shaping unit 2, the fluorescent signal receiving unit 6 and the 3 units of the 0-degree and 10-degree signal receiving units 4. Wherein the emitting end of the light source shaping unit 2 is connected with the incident end of the sheath flow device 3, one emitting end of the sheath flow device 3 is connected with the incident end of the 0 degree and 10 degree signal receiving unit 4, the other emitting end of the sheath flow device 3 is connected with the incident end and the emitting end of the light guiding unit 5, and the emitting end of the light guiding unit 5 is connected with the incident end of the fluorescent signal receiving unit 6. The laser beam outputted from the light source shaping means 2 is directly irradiated to the capillary holes in the sheath flow device 3. The laser beam, when irradiated on the cells in the capillary holes through the sheath flow device 3, generates a scattered light signal. A part of the scattered light signal directly enters the 0-degree and 10-degree signal receiving unit 4, and another part of the scattered light signal enters the fluorescent signal receiving unit 6 through the light guiding unit 5. The 0 degree and 10 degree signal receiving unit 4 and the fluorescent signal receiving unit 6 collect the light signals and send the light signals to the upper computer for cell analysis.
The light source shaping unit 2 mainly comprises 2 lasers 2-1, 4 cylindrical mirrors 2-2, 1 reflecting mirror 2-3, 1 light source dichroic mirror 2-4 and 1 light source focusing lens 2-5. The 2 lasers 2-1 have different wavelengths. In this embodiment, one of the lasers 2-1 is a 635nm red laser 2-1 and the other laser 2-1 is a 488nm blue laser 2-1. One of the lasers 2-1 passes through 2 mutually orthogonal cylindrical mirrors 2-2 and then enters a reflecting mirror 2-3, and is reflected by the reflecting mirror 2-3 and then enters one of the incident ends of a light source dichroic mirror 2-4. The other laser 2-1 passes through 2 mutually orthogonal cylindrical mirrors 2-2 and then directly enters the other incident end of the light source dichroic mirror 2-4. The outgoing end of the light source dichroic mirror 2-4 is incident into the capillary holes of the sheath flow device 3 via the light source focusing lens 2-5. After a circular laser spot emitted by a laser 2-1 passes through 2 mutually orthogonal cylindrical mirrors 2-2, the circular laser spot is compressed by the cylindrical mirrors 2-2 into an elliptical spot with a specific size, then the elliptical spot irradiates on a reflecting mirror 2-3, passes through a light source dichroic mirror 2-4 after being reflected by the reflecting mirror 2-3, and finally is focused on a sheath flow device 3 through a light source focusing lens 2-5 to perform particle detection. Meanwhile, after the circular laser spot emitted by the other laser 2-1 passes through 2 mutually orthogonal cylindrical mirrors 2-2, the circular laser spot is compressed into an elliptical spot with a specific size by the cylindrical mirrors 2-2, and then the elliptical spot directly passes through the light source dichroic mirror 2-4 and is focused on the sheath flow device 3 through the light source focusing lens 2-5 for particle detection. The cylindrical mirror 2-2 for spot shaping can be arranged outside the light outlet of the laser 2-1, and the cylindrical mirror 2-2 for spot shaping can be arranged inside the laser 2-1, namely the integrated shaping laser 2-1. See fig. 4.
The capillary holes of the sheath flow device 3 store cells to be measured. The sheath flow device 3 is arranged on the base 1 through an azimuth adjusting plate, and the azimuth adjusting plate can drive the sheath flow device 3 to realize the movement in the X-axis, Y-axis and Z-axis directions, so that the capillary holes of the sheath flow device 3 are ensured to be exactly positioned on the light path junction of the light source shaping unit 2, the fluorescent signal receiving unit 6 and the 3 units of the 0-degree and 10-degree signal receiving unit 4. When an elliptical light spot with a specific size emitted by the light source shaping unit 2 irradiates a particle to be detected in the sheath flow device 3, the light beam generates 360-degree light scattering along the particle and is received by the fluorescent signal receiving unit 6 and the 0-degree and 10-degree signal receiving units 4 respectively.
The 0-degree and 10-degree signal receiving unit 4 is composed of a receiving base 4-1, a diaphragm 4-2, a forward collecting lens 4-3, a perforated lens 4-4, a 0-degree signal receiving circuit board 4-5 and a 10-degree signal receiving circuit board 4-6. The receiving base 4-1 is internally provided with a light-in through hole seat 4-1-1, a 0-degree light-out through hole 4-1-2 and a 10-degree light-out through hole 4-1-3 which are communicated. The 0-degree light-emitting through hole 4-1-2 coincides with the central axis of the light-entering through hole seat 4-1-1, the 10-degree light-emitting through hole 4-1-3 is perpendicular to the central axis of the light-entering through hole seat 4-1-1, and the central axes of the light-entering through hole seat 4-1-1, the 0-degree light-emitting through hole 4-1-2 and the 10-degree light-emitting through hole 4-1-3 are intersected at the intersection of the 3 through holes. A perforated mirror 4-4 is arranged at this intersection. The outlet of the light inlet through hole seat 4-1-1 is connected with the inlet of the 0-degree light outlet through hole 4-1-2 and the inlet of the 10-degree light outlet through hole 4-1-3 through the perforated mirror 4-4. The entrance of the light entrance through hole seat 4-1-1 faces the sheath flow device 3. The forward condenser lens 4-3 is arranged at the entrance of the light-entering through hole seat 4-1-1. The diaphragm 4-2 is arranged at the front side of the front condenser 4-3, and mainly serves to block the light beam which does not diffract or scatter with particles, thereby improving the detection reliability and reducing the background voltage of the signal receiving circuit board. The 0-degree signal receiving circuit board 4-5 is arranged at the outlet of the 0-degree light emitting through hole 4-1-2. The 10-degree signal receiving circuit board 4-6 is arranged at the outlet of the 10-degree light emitting through hole 4-1-3. When the scattered light beam of the sheath flow device 3 passes through the forward collecting lens 4-3, the scattered light beam is gradually collected, a part of the light beam with small angle passes through the perforated lens 4-4 and propagates to the 0-degree signal receiving circuit board 4-5 through the 0-degree light outlet through hole 4-1-2, and another part of the light beam with large angle is reflected by the perforated lens 4-4 and propagates to the 10-degree signal receiving circuit board 4-6 through the 10-degree light outlet through hole 4-1-3, thereby completing the signal receiving of the optical signal in the 0-degree and 10-degree directions. See fig. 5.
When the laser beam irradiates the cell particles specifically bound to the fluorescent particles in the sheath flow device 3, fluorescence excitation is performed to generate stokes frequency domain, and the wavelength of the laser beam is caused to transit to the infrared direction. A light guide unit 5 is provided in the direction of advance of the light beam to guide the fluorescent signal into the fluorescent signal receiving unit 6. In the present embodiment, the light guiding unit 5 adopts a micro-collimating lens group, which mainly comprises a light guiding base 5-1, a front light guiding meniscus lens 5-2, a rear light guiding meniscus lens 5-3, a double-cemented lens 5-4, a micro-objective lens barrel 5-5, a slit baffle 5-7, a plano-concave micro-eyepiece 5-8 and a plano-convex micro-eyepiece 5-9. The light guide base 5-1 is internally provided with a light guide through hole 5-1-1. The entrance of the light guide through hole 5-1-1 faces the sheath flow device 3, and the exit of the light guide through hole 5-1-1 faces the fluorescent signal receiving unit 6. The front light guide meniscus lens 5-2, the rear light guide meniscus lens 5-3, the double-cemented lens 5-4 and the microscope objective lens are sequentially arranged at the entrance of the light guide through hole 5-1-1 from front to back, the front light guide meniscus lens 5-2 is arranged at one side close to the entrance of the light guide through hole 5-1-1, and the microscope objective lens is arranged at one side close to the exit of the light guide through hole 5-1-1. The slit baffle 5-7, the plano-concave microscopic ocular 5-8 and the plano-convex microscopic ocular 5-9 are sequentially arranged at the outlet of the light guide through hole 5-1-1 from front to back, the slit baffle 5-7 is arranged at one side close to the inlet of the light guide through hole 5-1-1, and the plano-convex microscopic ocular 5-9 is arranged at one side close to the outlet of the light guide through hole 5-1-1. The slit baffle 5-7 has the function of splitting the forward scattered light signal into two light components of 1-3 degrees and 7-11 degrees. The microscope objective lens barrel 5-5 is provided with a microscope objective adjusting piece 5-6 for adjusting the focus of the microscope objective lens barrel 5-5. The focal length of the microscopic imaging lens group is 35-100 mm. The fluorescent signal scattered by the sheath flow device 3 sequentially passes through a front light guide meniscus lens 5-2, a rear light guide meniscus lens 5-3, a double-cemented lens 5-4 and a microscope objective lens barrel 5-5 at the entrance of the light guide unit 5, then sequentially passes through a slit baffle 5-7, a plano-concave microscopic ocular 5-8 and a plano-convex microscopic ocular 5-9 at the entrance of the light guide unit 5, and finally is collimated and focused on a fluorescent signal receiving unit 6 at the rear end. See fig. 6.
The above-mentioned fluorescent signal receiving unit 6 is composed of a signal receiving housing 6-1, and 5 fluorescent dichroic mirrors 6-2, 6 bandpass filters 6-3, and 6 photoelectric converters 6-4 provided in the signal receiving housing 6-1. All the fluorescent dichroic mirrors 6-2 are removably mounted in the signal receiving housing 6-1 by means of a pluggable connection. The angle range of all the fluorescent dichroic mirrors 6-2 and the fluorescent scattering signal main light rays is 25-50 degrees. The signal receiving housing 6-1 is internally provided with light-absorbing cotton, which can reflect and transmit the fluorescent dichroic mirror 6-2 to be irradiated with the residual wavelength, thereby avoiding unnecessary photoelectric signal generation. The 5 fluorescent dichroic mirrors 6-2 have different transmission wavelengths, and the fluorescent dichroic mirrors 6-2 are installed in the signal receiving housing 6-1 at an angle of 45 degrees, and mainly function to separate light beams of different wavelengths, reflect back a specific wavelength in a 90 ° direction, and allow the specific wavelength to pass through. As the light passes stepwise through fluorescent dichroic mirror 6-2, the light intensity will decrease stepwise, while the particle information carried will decrease stepwise. The incident end of the first fluorescent dichroic mirror 6-2 forms the incident end of the fluorescent signal receiving unit 6. The 5 bandpass filters 6-3 have different filter wavelengths, and mainly function to block the passage of unspecific wavelengths and allow only specific wavelengths to pass, thereby functioning to purify signals. The band-pass filter 6-3 is arranged in the receiving shell by adopting a pluggable counter. The photoelectric converter 6-4 is mainly used for converting the optical signal into an electrical signal to be output and transmitting the electrical signal to the upper computer to display the characteristics of the particles carried by the electrical signal. In this embodiment, the photoelectric converter 6-4 may be a photomultiplier tube, an avalanche photodiode, a photodiode, or other charge coupled device.
The reflection output end of the first fluorescent dichroic mirror 6-2 is connected with the first photoelectric converter 6-4 via the first bandpass filter 6-3, and the excited fluorescent scattering signal chief ray is incident on the first fluorescent dichroic mirror 6-2 at an angle of 45 °. The transmission output end of the first fluorescent dichroic mirror 6-2 is connected to the incident end of the second fluorescent dichroic mirror 6-2. The reflective output end of the second fluorescent dichroic mirror 6-2 is connected to the incident end of the fifth fluorescent dichroic mirror 6-2, and the transmissive output end of the second fluorescent dichroic mirror 6-2 is connected to the incident end of the third fluorescent dichroic mirror 6-2. The reflection output end of the fifth fluorescent dichroic mirror 6-2 is connected to the sixth photoelectric converter 6-4 via a sixth bandpass filter 6-3. The transmission output end of the fifth fluorescent dichroic mirror 6-2 is connected to the fifth photoelectric converter 6-4 via a fifth bandpass filter 6-3. The incident output end of the third fluorescent dichroic mirror 6-2 is connected to the incident end of the fourth fluorescent dichroic mirror 6-2. The transmission output end of the third fluorescent dichroic mirror 6-2 is connected to the second photoelectric converter 6-4 via a second bandpass filter 6-3. The reflection output end of the fourth fluorescent dichroic mirror 6-2 is connected to the third photoelectric converter 6-4 via a third bandpass filter 6-3. The transmission output end of the fourth fluorescent dichroic mirror 6-2 is connected to the fourth photoelectric converter 6-4 via a fourth bandpass filter 6-3. See fig. 7 and 8.
Embodiment two:
the second embodiment is similar to the first embodiment in terms of the structure of the optical device, except that the light guiding unit 5 employs an optical fiber bundle 5-10 instead of a micro-collimating lens group, and one end of the optical fiber bundle 5-10 is coupled to the sheath fluid device 3, and the other end is coupled to the incident end of the fluorescent signal receiving unit 6. See fig. 9. The Y-shaped light transmission type optical fiber bundles 5-10 can realize micro-loss of optical signals, and the loss coefficient of a common standard single-mode optical fiber at 1550nm is 0.2dB/Km. The distance between the optical fiber signal receiving end and the sheath flow device 3 is 2-10 mm. In addition, a plano-convex lens with a large numerical aperture is additionally arranged between the optical fiber bundles 5-10 and the sheath fluid device 3, and the plano-convex lens has the main function of collecting light beams and better coupling the light beams into the optical fibers.
It should be noted that, although the examples described above are illustrative, this is not a limitation of the present invention, and thus the present invention is not limited to the above-described specific embodiments. Other embodiments, which are apparent to those skilled in the art from consideration of the specification and practice of the invention disclosed herein, are considered to be within the scope of the invention as claimed.
Claims (8)
1. An optical device for counting and detecting cells is characterized by mainly comprising a base (1), a light source shaping unit (2), a sheath flow device (3), 0-degree and 10-degree signal receiving units (4), a light guide unit (5) and a fluorescent signal receiving unit (6), wherein the light source shaping unit (2), the sheath flow device (3), the 0-degree and 10-degree signal receiving unit (4) and the fluorescent signal receiving unit (6) are arranged on the base (1); the sheath flow device (3) is positioned on the light path junction of the light source shaping unit (2), the fluorescent signal receiving unit (6) and the 0-degree and 10-degree signal receiving unit (4); the light source shaping device comprises a light source shaping unit (2), a sheath flow device (3), a signal receiving unit (4) and a fluorescent signal receiving unit (6), wherein the emitting end of the light source shaping unit (2) is connected with the incident end of the sheath flow device (3), one emitting end of the sheath flow device (3) is connected with the incident end of the signal receiving unit (4) with 0 degree and 10 degrees, the other emitting end of the sheath flow device (3) is connected with the incident end of the light guiding unit (5), and the emitting end of the light guiding unit (5) is connected with the incident end of the fluorescent signal receiving unit (6);
the 0-degree and 10-degree signal receiving unit (4) consists of a receiving base (4-1), a diaphragm (4-2), a forward collecting lens (4-3), a perforating lens (4-4), a 0-degree signal receiving circuit board (4-5) and a 10-degree signal receiving circuit board (4-6); the receiving base (4-1) is internally provided with a light-in through hole seat (4-1-1), a 0-degree light-out through hole (4-1-2) and a 10-degree light-out through hole (4-1-3) which are communicated with each other; the 0-degree light-emitting through hole (4-1-2) coincides with the central axis of the light-entering through hole seat (4-1-1), the 10-degree light-emitting through hole (4-1-3) is perpendicular to the central axis of the light-entering through hole seat (4-1-1), and the central axes of the light-entering through hole seat (4-1-1), the 0-degree light-emitting through hole (4-1-2) and the 10-degree light-emitting through hole (4-1-3) are intersected at the intersection of the 3 through holes; the perforated mirror (4-4) is arranged at the junction, and the outlet of the light inlet through hole seat (4-1-1) is connected with the inlet of the 0-degree light outlet through hole (4-1-2) and the inlet of the 10-degree light outlet through hole (4-1-3) through the perforated mirror (4-4); the entrance of the light-entering through hole seat (4-1-1) forms the incident end of the 0-degree and 10-degree signal receiving unit (4) and faces the sheath flow device (3), and the forward collecting lens (4-3) is arranged at the entrance of the light-entering through hole seat (4-1-1); the 0-degree signal receiving circuit board (4-5) is arranged at the outlet of the 0-degree light emitting through hole (4-1-2), and the 10-degree signal receiving circuit board (4-6) is arranged at the outlet of the 10-degree light emitting through hole (4-1-3);
the fluorescent signal receiving unit (6) consists of a signal receiving shell (6-1), and 5 fluorescent dichroic mirrors (6-2) and 6 photoelectric converters (6-4) which are arranged in the signal receiving shell (6-1); the incident end of the first fluorescent dichroic mirror (6-2) forms the incident end of the fluorescent signal receiving unit (6); the reflection output end of the first fluorescent dichroic mirror (6-2) is connected with the first photoelectric converter (6-4); the transmission output end of the first fluorescent dichroic mirror (6-2) is connected with the incident end of the second fluorescent dichroic mirror (6-2); the reflection output end of the second fluorescent dichroic mirror (6-2) is connected with the incidence end of the fifth fluorescent dichroic mirror (6-2), and the transmission output end of the second fluorescent dichroic mirror (6-2) is connected with the incidence end of the third fluorescent dichroic mirror (6-2); the reflection output end of the fifth fluorescent dichroic mirror (6-2) is connected with the sixth photoelectric converter (6-4); the transmission output end of the fifth fluorescent dichroic mirror (6-2) is connected with the fifth photoelectric converter (6-4); the incidence output end of the third fluorescent dichroic mirror (6-2) is connected with the incidence end of the fourth fluorescent dichroic mirror (6-2); the transmission output end of the third fluorescent dichroic mirror (6-2) is connected with the second photoelectric converter (6-4); the reflection output end of the fourth fluorescent dichroic mirror (6-2) is connected with the third photoelectric converter (6-4); the transmission output end of the fourth fluorescent dichroic mirror (6-2) is connected with the fourth photoelectric converter (6-4).
2. An optical device for cell counting and detection according to claim 1, characterized in that the output of the fluorescent dichroic mirror (6-2) is connected to the photoelectric converter (6-4) via a bandpass filter (6-3).
3. An optical device for cell counting and detection according to claim 1, characterized in that the fluorescent dichroic mirror (6-2) is removably mounted in the signal receiving housing (6-1) by means of a pluggable connection.
4. The optical device for cell counting and detection according to claim 1, wherein the light guiding unit (5) is composed of a light guiding base (5-1), a front light guiding meniscus lens (5-2), a rear light guiding meniscus lens (5-3), a double-cemented lens (5-4), a microscope objective lens barrel (5-5), a slit baffle (5-7), a plano-concave microscopic eyepiece (5-8) and a plano-convex microscopic eyepiece (5-9);
a light guide through hole (5-1-1) is arranged in the light guide base (5-1); the entrance of the light guide through hole (5-1-1) forms the incident end of the light guide unit (5) and faces the sheath flow device (3); the outlet of the light guide through hole (5-1-1) forms the emergent end of the light guide unit (5) and faces the fluorescent signal receiving unit (6); the front light guide meniscus lens (5-2), the rear light guide meniscus lens (5-3), the double-cemented lens (5-4) and the microscope objective lens are sequentially arranged at the entrance of the light guide through hole (5-1-1) from front to back, the front light guide meniscus lens (5-2) is arranged at one side close to the entrance of the light guide through hole (5-1-1), and the microscope objective lens is arranged at one side close to the exit of the light guide through hole (5-1-1); the slit baffle (5-7), the plano-concave microscopic ocular (5-8) and the plano-convex microscopic ocular (5-9) are sequentially arranged at the outlet of the light guide through hole (5-1-1) from front to back, the slit baffle (5-7) is arranged at one side close to the inlet of the light guide through hole (5-1-1), and the plano-convex microscopic ocular (5-9) is arranged at one side close to the outlet of the light guide through hole (5-1-1).
5. An optical device for cell counting and detection according to claim 1, characterized in that the light guiding unit (5) is a fiber bundle (5-10), one end of the fiber bundle (5-10) being coupled to the sheath fluid (3) and the other end being coupled to the entrance end of the fluorescent signal receiving unit (6).
6. An optical device for cell counting and detection according to claim 5, characterized in that a plano-convex lens is added between the optical fiber bundle (5-10) and the sheath flow device (3).
7. An optical device for cell counting and detection according to claim 1, characterized in that the light source shaping unit (2) mainly consists of 2 lasers (2-1), 4 cylindrical mirrors (2-2), 1 mirror (2-3), 1 light source dichroic mirror (2-4) and 1 light source focusing lens (2-5);
the 2 lasers (2-1) have different wavelengths, wherein one laser (2-1) passes through 2 mutually orthogonal cylindrical mirrors (2-2) and then enters a reflecting mirror (2-3), and is reflected by the reflecting mirror (2-3) and then enters one incident end of a light source dichroic mirror (2-4); the other laser (2-1) passes through 2 mutually orthogonal cylindrical mirrors (2-2) and then directly enters the other incident end of the light source dichroic mirror (2-4); the emergent end of the light source dichroic mirror (2-4) is connected with the incident end of the light source focusing lens (2-5), and the emergent end of the light source focusing lens (2-5) forms the emergent end of the light source shaping unit (2).
8. An optical device for counting and detecting cells according to claim 1, characterized in that the sheath flow device (3) is mounted on the base (1) by means of an orientation adjustment plate enabling adjustment in the X-axis, Y-axis and Z-axis directions.
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN109612911B (en) * | 2018-12-26 | 2023-07-21 | 深圳天依生命健康科技有限公司 | Full-automatic sperm cell detector |
| CN111117860B (en) * | 2019-11-04 | 2022-04-19 | 桂林电子科技大学 | Cell micro-scalpel based on optical fiber glimmer hand and preparation method thereof |
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