CN111889153A - Flow cytometer based on optical fiber integrated microfluidic chip - Google Patents
Flow cytometer based on optical fiber integrated microfluidic chip Download PDFInfo
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- 239000013307 optical fiber Substances 0.000 title claims abstract description 111
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- 239000012530 fluid Substances 0.000 claims abstract description 15
- 239000000835 fiber Substances 0.000 claims description 58
- 230000005540 biological transmission Effects 0.000 claims description 25
- 230000003287 optical effect Effects 0.000 claims description 17
- 238000012360 testing method Methods 0.000 claims description 5
- 238000000227 grinding Methods 0.000 claims description 4
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- 238000012216 screening Methods 0.000 claims description 3
- 238000004458 analytical method Methods 0.000 abstract description 6
- 238000000034 method Methods 0.000 abstract description 3
- 238000010586 diagram Methods 0.000 description 15
- 239000007788 liquid Substances 0.000 description 4
- 230000000694 effects Effects 0.000 description 3
- 238000000684 flow cytometry Methods 0.000 description 3
- 230000001413 cellular effect Effects 0.000 description 2
- 238000013461 design Methods 0.000 description 2
- 238000004720 dielectrophoresis Methods 0.000 description 2
- 238000001914 filtration Methods 0.000 description 2
- 230000010354 integration Effects 0.000 description 2
- 238000005259 measurement Methods 0.000 description 2
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- B01L3/5027—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures by integrated microfluidic structures, i.e. dimensions of channels and chambers are such that surface tension forces are important, e.g. lab-on-a-chip
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- G01N15/10—Investigating individual particles
- G01N15/14—Optical investigation techniques, e.g. flow cytometry
- G01N15/1434—Optical arrangements
- G01N15/1436—Optical arrangements the optical arrangement forming an integrated apparatus with the sample container, e.g. a flow cell
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Abstract
The invention provides a flow cytometer based on an optical fiber integrated microfluidic chip. The method is characterized in that: the device consists of a micro-fluidic control system 1, an optical fiber integrated micro-fluidic chip 2, an excitation light source 3 and a scattered light detection module 4. In the system, a micro-fluidic system 1 controls the flow rate of a cell sample and sheath fluid flowing into a micro-fluidic chip 2, a single cell flow is formed in a single cell flow channel 2-6, optical fibers are embedded on the micro-fluidic chip 2 and distributed on two sides of the micro-fluidic channel, a first optical fiber 2-1 is used for inputting an excitation light beam, and a second optical fiber 2-2 and a third optical fiber 2-3 are used for collecting scattered light. Scattered signal light in different directions is collected through optical fibers and transmitted to the scattered light detection module 4 for light splitting detection. The invention can be used for automatic analysis and sorting of cells.
Description
(I) technical field
The invention relates to a flow cytometer based on an optical fiber integrated microfluidic chip, which can be used for automatic analysis and sorting of cells and belongs to the technical field of cell detection and analysis.
(II) background of the invention
Flow cytometry, a cellular analysis technique, was first used to measure cell volume in the 50's 20 th century and can detect cells as they pass straight through a viewing hole with a fast flowing fluid. In flow cytometry, cells in solution are detected by passing the laser beam of the instrument at a rate of 10,000 cells (or more) per second. Today's flow cytometers have more detectable fluorescence parameters (from 1 or 2 up to about 30 or more) and can measure all these parameters on the same cell simultaneously. Flow cytometry is fast and has single cell level detection capability, can provide statistical functionality for cell biologists, and can rapidly analyze and characterize millions of cells.
The flow cytometer mainly comprises a fluid system, an optical system and a photoelectric processing system. The fluidic system is a life-line of a flow cytometer. It is responsible for aligning the cells in the core stream in a single column and passing them to the checkpoint for data collection. If sample introduction is inconsistent and cell alignment is incorrect, this can result in significant data diffusion, thereby reducing the confidence in the data. The traditional fluid system mainly adopts a nozzle (also called a fluid chamber) to spray a single cell flow which is formed by sheath fluid and sample flow together, then space laser irradiates on the single cell flow, forward scattered light and lateral scattered light are collected and dispersed, and then the single cell flow is processed and analyzed by a photoelectric detection system to obtain the statistical data of cells.
The traditional flow cytometer adopts a spatial light path, has high requirement on the stability of the light path, needs to precisely adjust the light path, and has large interference of external environments such as vibration, temperature, humidity and the like on optical elements. Moreover, these space optical elements are bulky and are not flexible enough in assembly.
Disclosure of the invention
The invention aims to provide a flow cytometer based on an optical fiber integrated microfluidic chip.
The purpose of the invention is realized as follows:
as shown in fig. 1, a flow cytometer based on an optical fiber integrated microfluidic chip. The device consists of a micro-fluidic control system 1, an optical fiber integrated micro-fluidic chip 2, an excitation light source 3 and a scattered light detection module 4. In the system, a micro-fluidic system 1 controls the flow rate of a cell sample and sheath fluid flowing into a micro-fluidic chip 2, a single cell flow is formed in a single cell flow channel 2-6, optical fibers are embedded on the micro-fluidic chip 2 and distributed on two sides of the micro-fluidic channel, a first optical fiber 2-1 is used for inputting an excitation light beam, and a second optical fiber 2-2 and a third optical fiber 2-3 are used for collecting scattered light. Scattered signal light in different directions is collected through optical fibers and transmitted to the scattered light detection module 4 for light splitting detection.
The optical fiber integrated microfluidic chip 2 is provided with two sheath fluid inflow channels 2-4, a cell sample flows into the channels 2-5, and the flow velocity of each channel is controlled by a microfluidic control system to form single cell flow; an excitation light transmission fiber 2-1 and scattered light collection fibers 2-2 and 2-3 are fixed on two sides of the single-cell flow channel 2-6; the tail end of the single-cell flow channel 2-6 is connected with a cell sorting channel 2-7, and microelectrodes 2-8 are embedded at two sides for screening and classifying cells.
Alternatively, as shown in fig. 2, the microfluidic chip 2 may be a first type microfluidic chip. The optical fibers on the microfluidic chip are integrated in the plane of the microfluidic chip 2, wherein the exciting light transmission optical fiber 2-1 is vertical to the direction of the single-cell flow channel 2-6. The scattered light collection optical fiber 2-2 is a forward scattered light collection optical fiber and is arranged opposite to the exciting light transmission optical fiber 2-1; the scattered light collection fibers 2-3 are side scattered light collection fibers and the forward scattered light collection fibers 2-2 are arranged at an angle α, 0< α <180 ° and α ≠ 90 °.
Alternatively, as shown in fig. 3, the microfluidic chip 2 may be a second type microfluidic chip. The exciting light transmission optical fiber 2-1 and the forward scattering light collection optical fiber 2-2 of the microfluidic chip are embedded in the plane of the microfluidic chip and are oppositely arranged on two sides of the single-cell flow channel 2-6, and the lateral scattering light collection optical fiber 2-3, the exciting light transmission optical fiber 2-1 and the forward scattering light collection optical fiber 2-2 are coplanar and perpendicular to the microfluidic chip 2, and lateral scattering light is collected from the bottom of the single-cell flow channel 2-6.
Alternatively, as shown in fig. 4, the microfluidic chip 2 may be a third type microfluidic chip. The optical fibers of the microfluidic chip are integrated in the plane of the microfluidic chip 2. The exciting light transmission fiber 2-1 and the forward scattered light collection fiber 2-2 are oppositely arranged on two sides of the single-cell flow channel 2-6. The angle between the excitation light delivery fiber 2-1 and the single-cell flow channel 2-6 is β, 0< β <90 °, preferably β ═ 45 °. The side scattered light collecting fiber 2-3 is disposed perpendicularly to the forward scattered light collecting fiber 2-2. The number of the side scattered light collecting fibers 2 to 3 may be one (fig. 4) or two (fig. 5). Multiple side scatter collection fibers may be used to obtain more abundant fluorescence information.
Optionally, as shown in fig. 6, a fourth type of microfluidic chip is provided, and the excitation light transmission fiber 2-1, the forward scattered light collection fiber 2-2, and the lateral scattered light collection fiber 2-3 are used as a set of optical path testing system of the flow cytometer, and a plurality of optical path testing systems can be integrated on the same microfluidic chip. This is similar to the parallel light path excitation device of the traditional flow cytometer, i.e. single cells can pass light beams with different wavelengths at different positions, thus leading to richer fluorescence detection information. For example, if the excitation wavelength bands of the two fluorescent markers are different but the emission wavelengths are the same, the wavelengths of the fluorescence signals obtained by common-path excitation are the same, and thus the two markers cannot be distinguished. And the parallel light path separates the two excitation wavelengths to excite at different positions and detect the two excitation wavelengths respectively, so that the markers can be distinguished.
Optionally, the excitation light transmission fiber may be one of a single mode fiber, a multi-core fiber or a ring-core fiber; the end face of the optical fiber may be a flat end obtained by cutting or a focusing lens end obtained by grinding.
Alternatively, the excitation light source may be a single-wavelength laser light source or multiple light sources with different wavelengths, and the excitation light is coupled into one excitation light transmission fiber via a fiber wavelength division multiplexer for transmission, which is similar to a common-path excitation system in a conventional flow cytometer, that is, multiple wavelengths of laser light simultaneously act on each cell passing through the light beam.
The scattered light detection module is of an optical fiber type. The forward scattered light is transmitted to a photoelectric detector through a forward scattered light collection optical fiber, and a band-pass filter film is plated on the end face of the tail end of the optical fiber to filter signal light with other wavelengths by exciting the forward scattered light with the wavelength; after the side scattered light is collected by the side scattered light collecting optical fibers, the side scattered light is divided into multiple paths of signal light by the broadband coupler and is respectively transmitted to different photoelectric detectors for detection, and different band-pass filter coatings are plated at the tail end of each optical fiber and penetrate through scattered light and fluorescent signals with different wavelengths.
The scattered light detection module is of a space optical module type. The forward scattered light is transmitted to a photoelectric detector for detection through a band-pass filter; the side scattered light passes through a plurality of dichroic mirrors, and the fluorescence is sorted to different photoelectric detectors for detection.
Compared with the traditional flow cytometer, the invention has the following advantages:
(1) compared with the traditional nozzle type microfluidic channel, the microfluidic chip has the advantages of various preparation methods, various material types, rich design diversity of the fluid channel, high integration level of the chip, small dimension and high design flexibility.
(2) The optical fiber is adopted to replace the traditional space optical path, so that the fine adjustment of the space optical path when a device is replaced is avoided. The optical fiber is directly embedded into the microfluidic chip, the chip can be integrally replaced, and the whole system is high in stability and is slightly influenced by external environments such as temperature, humidity and vibration. And the optical fiber has excellent flexibility and can be bent arbitrarily, which is extremely advantageous for integration and miniaturization of the system.
(3) The microfluidic chip can be used for analyzing cells with different sizes by replacing microfluidic channels with different sizes. For tiny cells, the micro-lens at the end of the optical fiber realizes the output of a light beam with a micron size, and ensures the measurement resolution when the cells pass through.
(IV) description of the drawings
Fig. 1 is a system diagram of a flow cytometer based on a fiber-optic integrated microfluidic chip.
Fig. 2 is a schematic structural diagram of a first type of optical fiber integrated microfluidic chip.
Fig. 3 is a schematic structural diagram of a microfluidic chip integrated with a second type of optical fiber.
Fig. 4 is a schematic structural diagram of a microfluidic chip integrated with a third type of optical fiber.
Fig. 5 is an extension of a microfluidic chip integrated with a third type of optical fiber, which has one more side scattered light collecting optical fiber 2-3 than the microfluidic chip of fig. 4.
Fig. 6 is a schematic structural diagram of a microfluidic chip integrated with a fourth type of optical fiber. Multiple optical path test systems can be integrated on the same microfluidic chip.
Fig. 7 is a diagram of an optical fiber end integrated type optical filtering system, which employs a method of coating an end surface of an optical fiber connector to implement narrow-band bandpass filtering, wherein (a) is a schematic position diagram of a coated dielectric film, (b) a spectrum of an optical signal collected by a scattered light collection optical fiber, and (c) a monochromatic spectrum obtained after passing through the dielectric film.
Fig. 8 is a structural diagram of a fiber-optic scattered light detection module of a flow cytometer based on a fiber-optic integrated microfluidic chip.
Fig. 9 is a structural view of a spatial light module type scattered light detection module.
Fig. 10 is a diagram of a flow cytometer system based on an optical fiber integrated microfluidic chip using spatial light modular scatter detection.
FIG. 11 is a diagram of an end face structure of a ring core optical fiber.
Fig. 12 is a schematic structural diagram of an excitation light path module formed by inputting a multi-wavelength excitation light source into an annular core optical fiber.
FIG. 13 shows microlens structures for focusing the ring beam at the end of the ring core fiber, (a) a symmetrical truncated cone structure obtained by fine grinding, and (b) an optimized curved truncated cone structure.
FIG. 14(a) is a 3D schematic diagram of the cone-frustum structure of the end face of the ring core fiber for focusing the light beam, and (b) is an axial sectional view showing the high-resolution excitation effect of the cone-frustum structure on the cell in the microfluidic chip.
(V) detailed description of the preferred embodiments
The invention is further illustrated with reference to the following figures and specific examples.
Example 1:
fig. 1 is a system diagram of a flow cytometer based on an optical fiber integrated microfluidic chip. The device consists of a micro-fluidic control system 1, an optical fiber integrated micro-fluidic chip 2, an excitation light source 3 and a scattered light detection 4. A micro-flow controller 1-1 in a micro-flow control system 1 in the system controls the flow velocity of a cell sample and sheath fluid flowing into a micro-flow chip through a guide pipe 1-2, a single cell flow is formed on an optical fiber integrated micro-flow chip 2, and optical fibers perpendicular to the direction of a single cell flow channel 2-6 are embedded on the micro-flow chip 2 and distributed on two sides of the micro-flow channel. One side of the optical fiber is an excitation light transmission optical fiber 2-1 which is used for inputting an excitation light source 3 light beam; the other side of the optical fiber is a scattered light collecting optical fiber 2-2/2-3 for collecting scattered light. Scattered signal light in different directions is collected through optical fibers 2-2/2-3 and transmitted to a scattered light detection module 4 for light splitting and detection.
Fig. 2 is a schematic structural diagram of a first type of optical fiber integrated microfluidic chip. The microfluidic chip is provided with a cell liquid and sheath liquid inlet pool 2-0, two sheath liquid inlet channels 2-4 and a cell sample inlet channel 2-5, and the flow velocity of each channel is controlled by a microfluidic control system 1 to form a single cell flow; micro-grooves are carved on two sides of the single-cell flow channel 2-6, and an excitation light transmission optical fiber 2-1 and a scattered light collection optical fiber 2-2/2-3 are fixed; the end of the single-cell flow channel 2-6 is connected with a cell sorting channel 2-7, the width of the channel is increased to two times of that of the single-cell flow channel 2-6, the aim is to reduce the flow rate of the cell flow, microelectrodes 2-8 are embedded on two sides of the cell sorting channel 2-7 and used for cell screening, different cells can generate different dielectrophoresis forces and deflect the cells, and therefore different types of cells can flow into different cell pools 2-9.
The radius of the liquid inlet pool 2-0 and the cell sap storage pool 2-9 of the optical fiber integrated microfluidic chip is R1mm, and preferably, R1 is 0.5.
The widths of the sheath fluid inflow channel 2-4, the cell sample inflow channel 2-5, and the single flow cell channel 2-6 are W1 μm. Preferably, W1 is 200.
All channels have a height of H μm, preferably H is 70.
The tail end of the single-cell flow channel is connected with a cell sorting channel 2-7, the width of the cell sorting channel 2-7 is W2 μm, which is twice of that of the single-cell flow channel 2-6, and the cell sorting channel is used for reducing the flow velocity of the single-cell flow and increasing the sorting efficiency. Preferably, W2 is 400.
The cell sorting channels 2-7 are flanked by microelectrodes 2-8, the purpose of which is to produce dielectrophoresis. The cells of the single-cell flow channel 2-7 can be sorted due to differences in cellular composition, membrane and size, and different dielectrophoretic forces generated by the presence of different polarization factors between different cells. By optimizing key parameters such as cell fluid flow velocity, sheath fluid pressure, voltage and frequency of the microelectrodes 2-8 and the like in the micro-fluidic system 1, cells can be effectively sorted at high speed and high efficiency.
The scattered light collection optical fiber comprises an optical fiber 2-2 which is right opposite to the exciting light transmission optical fiber and is used for collecting forward scattered light, and the size of the forward scattered light can reflect the size of the cell; one or more optical fibers 2-3 forming an acute angle with the direction of the single-cell flow channel are also included for collecting the side scattered light, and the more complicated the internal structure of the cell, the stronger the intensity of the side scattered light. Of course, the side scattered light also contains a large amount of fluorescent signals, and the signals need to be filtered and detected by filters with different parameters, so that the information of cell analysis parameters is increased.
The scattered light detection module 4 shown in fig. 1 comprises two parts: forward scatter detection and side scatter detection. The scattered light is collected by scattered light collection fibers 2-2/2-3 and transmitted to photodetector 4-1 for sensing. It is noted that, in order to distinguish fluorescence information of different wavelengths, a dielectric film 5-1 with different parameters is plated on the end face of each optical fiber adapter 5 connected to the photodetector, as shown in fig. 7 (a). The dielectric film 5-1 mainly functions as a narrow band pass filter, the scattered light collected by the scattered light collection fiber contains optical information of various wavelengths, as shown in fig. 3(b), and only optical information of a single wavelength is left after passing through the dielectric film and enters the photodetector for analysis, as shown in fig. 3 (c).
As shown in fig. 8, in order to separate different fluorescence signals from the side scattered light, the collected side scattered light needs to be split by a broadband fiber splitter 4-2, and then the fluorescence signals are detected by different photodetectors 4-1.
Example 2:
the optical fiber type scattered light detection module used in embodiment 1 needs to be coated with different dielectric films on the end face of the adapter at the tail end of the optical fiber due to the light splitting requirement. We can also use a filtered detection module based on spatial light paths. As shown in FIG. 10, the module is compact in composition and mainly comprises an adapter flange 6-1 for connecting a scattered light collection optical fiber, a dichroic mirror 6-2 with different parameters, a filter 6-3 with different parameters and a photoelectric detector 6-4. As shown in FIG. 6, the laterally scattered light passes through the lightThe fibers 2-3 are collected and transmitted to a scattered light detection module 6, and are divided into different wavelengths to be detected after passing through a plurality of dichroic mirrors 6-2 and optical filters 6-3. Wherein λ0Represents a Rayleigh scattered signal, λ, of the same wavelength as the excitation light1、λ2、λ3Respectively representing fluorescent signals of different wavelengths.
Example 3:
in both embodiments, either the excitation light transmitting fiber 2-1 or the scattered light collecting fiber 2-2/2-3 can be a single mode fiber. However, in order to compress the output beam of the excitation light transmission fiber, realize an output beam waist of several microns, and ensure the resolution of measurement when the cell passes through, a special fiber can be adopted and a focusing lens can be added at the end of the fiber.
This embodiment can employ a ring-core optical fiber 7 as shown in fig. 11, which is connected in a manner as shown in fig. 12. The laser of the excitation light sources 3-1 and 3-2 with two different wavelengths is transmitted by a single mode fiber 9 after being combined by a fiber wavelength division multiplexer 8. The fiber core of the single-mode fiber 9 is welded at a point aligned with the fiber core of the annular core fiber 7, so that the light beam is coupled into the annular core fiber 7, a frustum focusing structure 7-1 is prepared on the end face of the annular core fiber 7, and the reflection focusing of the excitation light beam transmitted in the annular core can be realized. The frustum focusing structure 7-1 is a symmetrical frustum cone structure obtained by fine grinding, as shown in fig. 13 (a). In order to achieve better beam focusing and higher resolution focused beam, an optimized curved frustum structure 7-2 can be used, as shown in fig. 13 (b).
FIG. 14(a) is a 3D schematic diagram of the focusing effect of the end-face frustum structure 7-1 of the ring-core optical fiber 7 on the light beam 10, and (b) shows that in the microfluidic chip, the focusing light beam is just in the middle of the single-cell flow channel 2-6, and single cells 11 pass through the focusing light beam one by one, so that the high-resolution excitation effect on the cells is realized.
Claims (10)
1. A flow cytometer based on an optical fiber integrated microfluidic chip is characterized in that: the device consists of a micro-fluidic control system 1, an optical fiber integrated micro-fluidic chip 2, an excitation light source 3 and a scattered light detection module 4. In the system, a micro-fluidic system 1 controls the flow rate of a cell sample and sheath fluid flowing into a micro-fluidic chip 2, a single cell flow is formed in a single cell flow channel 2-6, optical fibers are embedded on the micro-fluidic chip 2 and distributed on two sides of the micro-fluidic channel, a first optical fiber 2-1 is used for inputting an excitation light beam, and a second optical fiber 2-2 and a third optical fiber 2-3 are used for collecting scattered light. Scattered signal light in different directions is collected through optical fibers and transmitted to the scattered light detection module 4 for light splitting detection.
2. The flow cytometer based on the fiber integrated microfluidic chip as claimed in claim 1, wherein: the optical fiber integrated microfluidic chip 2 is provided with two sheath fluid inflow channels 2-4, a cell sample flows into the channels 2-5, and the flow velocity of each channel is controlled by a microfluidic control system to form single cell flow; an excitation light transmission fiber 2-1 and scattered light collection fibers 2-2 and 2-3 are fixed on two sides of the single-cell flow channel 2-6; the tail end of the single-cell flow channel 2-6 is connected with a cell sorting channel 2-7, and microelectrodes 2-8 are embedded at two sides for screening and classifying cells.
3. A flow cytometer based on an optical fiber integrated microfluidic chip as described in claim 1 and claim 2, wherein: the optical fibers are all integrated in the plane of the microfluidic chip 2, wherein the exciting light transmission optical fiber 2-1 is vertical to the direction of the single-cell flow channel 2-6. The scattered light collection optical fiber 2-2 is a forward scattered light collection optical fiber and is arranged opposite to the exciting light transmission optical fiber 2-1; the scattered light collection fibers 2-3 are side scattered light collection fibers and the forward scattered light collection fibers 2-2 are arranged at an angle α, 0< α <180 ° and α ≠ 90 °.
4. A flow cytometer based on an optical fiber integrated microfluidic chip as described in claim 1 and claim 2, wherein: the exciting light transmission optical fiber and the forward scattering light collection optical fiber are embedded in the surface of the microfluidic chip and are oppositely arranged on two sides of the single-cell flow channel, and the lateral scattering light collection optical fiber is coplanar with the exciting light transmission optical fiber and the forward scattering light collection optical fiber and is perpendicular to the microfluidic chip to collect lateral scattering light from the bottom of the single-cell flow channel.
5. A flow cytometer based on an optical fiber integrated microfluidic chip as described in claim 1 and claim 2, wherein: the optical fibers are all integrated in the plane of the microfluidic chip 2. The exciting light transmission optical fiber and the forward scattering light collection optical fiber are oppositely arranged at two sides of the single-cell flow channel. The included angle between the exciting light transmission fiber and the single-cell flow channel is beta, and 0< beta <90 degrees. The side scattered light collection fiber is positioned perpendicular to the forward scattered light collection fiber. The number of the side scattered light collecting fibers may be one or two.
6. A flow cytometer based on fiber-optic integrated microfluidic chip as described in claims 1, 2, 3, 4 and 5, wherein: the exciting light transmission optical fiber, the forward scattering light collection optical fiber and the lateral scattering light collection optical fiber are used as a set of flow cytometer optical path test system, and a plurality of same optical path test systems can be integrated on the same microfluidic chip.
7. A flow cytometer based on an optical fiber integrated microfluidic chip as described in claim 1 and claim 2, wherein: the exciting light transmission fiber can be one of a single-mode fiber, a multi-core fiber or a ring-core fiber; the end face of the optical fiber may be a flat end obtained by cutting or a focusing lens end obtained by grinding.
8. The flow cytometer based on the fiber integrated microfluidic chip as claimed in claim 1, wherein: the excitation light source can be a single-wavelength laser light source or a plurality of light sources with different wavelengths, and is coupled into the excitation light transmission optical fiber through the optical fiber wavelength division multiplexer for transmission.
9. The flow cytometer based on the fiber integrated microfluidic chip as claimed in claim 1, wherein: the scattered light detection module is of an optical fiber type. The forward scattered light is transmitted to a photoelectric detector through a forward scattered light collection optical fiber, and a band-pass filter film is plated on the end face of the tail end of the optical fiber to filter signal light with other wavelengths by exciting the forward scattered light with the wavelength; after the side scattered light is collected by the side scattered light collecting optical fibers, the side scattered light is divided into multiple paths of signal light by the broadband coupler and is respectively transmitted to different photoelectric detectors for detection, and different band-pass filter coatings are plated at the tail end of each optical fiber and penetrate through scattered light and fluorescent signals with different wavelengths.
10. The flow cytometer based on the fiber integrated microfluidic chip as claimed in claim 1, wherein: the scattered light detection module is of a space optical module type. The forward scattered light is transmitted to a photoelectric detector for detection through a band-pass filter; the side scattered light passes through a plurality of dichroic mirrors, and the fluorescence is sorted to different photoelectric detectors for detection.
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| CN112697679A (en) * | 2020-12-04 | 2021-04-23 | 杭州娃哈哈精密机械有限公司 | Device for rapidly detecting number of bacteria in beverage |
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