CN106383082B - A kind of optical path adjustment device and method of the flow cytometer without fluid path situation - Google Patents

Abstract

The invention provides an optical path adjustment device for flow cytometer without a liquid path, the device includes: an irradiation light source, an irradiation spot shaping optical path, a standard microsphere rotating device, a non-forward scattered light beam shaping optical path, a non-forward Scattered light detection circuit, forward scattered light and fluorescence beam shaping optical path, forward scattered light detection circuit, multicolor fluorescence spectroscopic optical path and multi-channel fluorescence detection circuit, wherein, the irradiation light source provides an excitation light source for fluorescence excitation; the irradiation The spot shaping optical path is used to compress the light source beam into an illumination spot of a certain size; the standard microsphere rotating device is used to rotate the disc loaded with standard microspheres to simulate single cells passing through the irradiation spot one by one; the non-forward The scattered light beam shaping optical path is used to focus the non-forward scattered light beam within a certain range; the non-forward scattered light detection circuit is used for photoelectric conversion of the non-forward scattered light and realizes parameterization of the generated electrical pulse signal extract.

Description

Optical path adjustment device and method for flow cytometer without liquid path

technical field

The invention relates to the field of optical path adjustment and microsphere measurement of a flow cytometer, in particular to an optical path adjustment device and method without a liquid path.

Background technique

In the flow cytometer optical path adjustment system, it includes the adjustment of the illumination spot and the spectroscopic collection of the excitation light signal. Since the detection area of the microspheres in the flow cytometer is located at the intersection of the irradiation spot and the direction of the liquid flow, and the instability of the liquid system will cause differences in the relative positions of the microspheres entering the detection area, resulting in the microspheres in the detection area The excitation degree of the irradiation is not the same, resulting in large differences in the intensity of forward scattered light, non-forward scattered light and fluorescence signal of each channel. The pulse parameter information obtained by the subsequent detection circuit is inaccurate. The existing optical adjustment system completely depends on the high-precision liquid circuit control system, which has poor independence. The hydraulic control system is extremely complex and cumbersome, and there is no standard index for the verification method of control accuracy and laminar flow effect.

Therefore, there is a need for an optical path adjustment device and method for a flow cytometer in the absence of a liquid path that can effectively solve the above problems.

Contents of the invention

According to one aspect of the present invention, a device for adjusting the optical path of a flow cytometer without a liquid path is provided. Scattered light beam shaping optical path, non-forward scattered light detection circuit, forward scattered light and fluorescence beam shaping optical path, forward scattered light detection circuit, multicolor fluorescence splitting optical path and multi-channel fluorescence detection circuit, wherein,

The irradiation light source provides an excitation light source for fluorescence excitation;

The illumination spot shaping optical path is used to compress the light source beam into an illumination spot of a certain size;

The standard microsphere rotating device is used to rotate the disc loaded with standard microspheres so as to simulate single cells passing through the irradiation spot one by one;

The non-forward scattered light beam shaping optical path is used to focus the non-forward scattered light beam within a certain range;

The non-forward scattered light detection circuit is used to photoelectrically convert the non-forward scattered light and realize parameter extraction for the generated electric pulse signal;

The forward scattered light and fluorescent beam shaping optical path is used to focus the forward scattered light and fluorescent beam within a certain range;

The forward scattered light detection circuit is used to photoelectrically convert the forward scattered light and extract parameters from the generated electric pulse signal;

The multi-color fluorescence spectroscopic optical path is used to separate the fluorescence signals in various wavelength ranges and transmit them to corresponding detection circuits. The multi-channel fluorescence detection circuit is used to perform photoelectric conversion on the fluorescence signals of each channel and realize Parameter extraction.

Preferably, a laser with a certain power and wavelength is used as the irradiation light source, and the laser emission beam is compressed and shaped through the irradiation spot shaping optical path. The irradiation spot shaping optical path realizes two orthogonal directions according to the movement direction of the microsphere in the standard microsphere rotating device. The spot size is compressed, and the moving microspheres are irradiated at the focal point of the spot by adjusting the light path of the spot shaping.

Preferably, the standard microsphere rotating device comprises a turntable loaded with standard microspheres, a turntable drive device and a motor motion control circuit, and the turntable loaded with standard microspheres is connected with the motor motion control circuit through the turntable drive device, through Adjusting the rotation speed of the motor realizes the control of the rotation speed of the turntable, thereby changing the irradiation time of the microspheres by the light spot and the time interval of the irradiation of adjacent microspheres.

Preferably, the turntable loaded with standard microspheres consists of a bottom layer and a top layer, wherein there are pits for placing standard microspheres in one layer, and the standard microspheres are sealed in the pits by bonding the top layer and the bottom layer to prevent microspheres. The ball falls off during the spin. During the rotation of the turntable loaded with standard microspheres, the standard microspheres pass through the irradiated spot area one by one to form side scattered light, forward scattered light and fluorescent signals of various colors.

Preferably, the non-forward scattered light beam shaping optical path is placed in the non-forward scattered light angle detection area, focusing and shaping the non-forward scattered light within a certain angle range, and transmitting the shaped beam to the non-forward scattered light detection circuit. The non-forward scattered light detection circuit realizes the photoelectric conversion of the non-forward scattered light signal and the conditioning processing of the electric pulse signal, and realizes the extraction of the electric pulse parameters characterizing the characteristics of the microsphere.

Preferably, the forward scattered light and fluorescent beam shaping optical path is placed in the direction of the irradiation beam, focusing and shaping the forward scattered light and multi-channel fluorescent beam within a certain angle range, and transmitting the shaped beam to the forward scattered light A detection circuit and a multi-color fluorescence spectroscopic light path. The forward scattered light detection circuit realizes the photoelectric conversion of the forward scattered light signal and the conditioning processing of the electric pulse signal, and realizes the extraction of the electric pulse parameters characterizing the characteristics of the microsphere.

Preferably, the multi-color fluorescence light splitting path separates multi-channel fluorescence signals through a series of optical devices, and transmits the separated fluorescent light beams of various colors to corresponding multi-channel fluorescence detection circuits. The multi-channel fluorescence detection circuit realizes the photoelectric conversion of the fluorescence signal and the conditioning processing of the electric pulse signal, and realizes the extraction of the electric pulse parameters characterizing the characteristics of the microsphere.

According to another aspect of the present invention, a method for adjusting the optical path of a flow cytometer without a liquid path is provided, comprising steps:

The irradiation spot shaping optical path compresses the light beam generated by the irradiation light source to obtain an irradiation spot of a certain size, and adjusts the focal length of the spot to make the microsphere pass through the focal point;

The standard microsphere rotating device adjusts the movement speed of the microsphere, changes the detection frequency of the microsphere and the irradiation time of the microsphere, thereby changing the frequency and duration of the corresponding light pulse and electric pulse;

The non-forward scattered light beam shaping optical path performs shaping and other processing on the non-forward scattered light, and adjusts the focal length of the optical path so that the detector in the non-forward scattered light detection circuit is at the focal point;

The non-forward scattered light detection circuit realizes the photoelectric conversion, electric pulse conditioning, analog/digital conversion and parameter extraction of non-forward scattered light, and identifies the starting point and ending point of the electric pulse;

The forward scattered light and fluorescent beam shaping optical path performs shaping and other processing on the forward scattered light and fluorescent beam, and transmits the focused beam to the multicolor fluorescent beam splitting optical path;

The multi-color fluorescence spectroscopic optical path splits the forward scattered light and the fluorescent beams of each channel according to the wavelength information, and transmits the split beams to the forward scattered light detection circuit and the corresponding multi-channel fluorescence detection circuit;

The forward scattered light detection circuit realizes the photoelectric conversion, electric pulse conditioning, analog/digital conversion and parameter extraction of forward scattered light, and identifies the starting point and ending point of the electric pulse;

The multi-channel fluorescence detection circuit realizes the photoelectric conversion, electric pulse conditioning, analog/digital conversion and parameter extraction of each fluorescence, and identifies the starting point and ending point of the electric pulse.

It should be understood that both the foregoing general description and the following detailed description are exemplary illustrations and explanations, and should not be used as limitations on the claimed content of the present invention.

Description of drawings

With reference to the accompanying drawings, more objects, functions and advantages of the present invention will be clarified through the following description of the embodiments of the present invention, wherein:

1 is a schematic diagram of an embodiment of an optical path adjustment device for a flow cytometer of the present invention without a liquid path;

Fig. 2 is a schematic diagram of the composition and connection of the flow cytometer of the present invention.

3a-3c are schematic diagrams of an exemplary structure of the illumination light source and the illumination spot shaping optical path shown in FIG. 2 . Fig. 3a shows the shape change of the light spot after the laser light emitted by the irradiation light source passes through the shaping lens. 3b-3c are schematic diagrams of light intensity distribution of the illuminated spot after shaping shown in FIG. 3a.

Figures 4a-4b show a schematic diagram of an implementation of a standard microsphere rotating device and standard microspheres.

Fig. 5 shows a structural diagram of a specific embodiment of an optical path of a schematic non-forward scattered light beam shaping optical path.

FIG. 6 shows a specific circuit block diagram of the non-forward scattered light detection circuit.

FIG. 7 shows a specific circuit block diagram of the forward scattered light detection circuit.

Fig. 8 shows a structural diagram of a specific optical path embodiment of a schematic multicolor fluorescence beam splitting optical path.

FIG. 9 shows a structural diagram of another schematic optical path embodiment of a multicolor fluorescence splitting optical path.

Fig. 10 shows a specific circuit block diagram of the multi-channel fluorescence detection circuit.

Detailed ways

Hereinafter, embodiments of the present invention will be described with reference to the accompanying drawings. In the drawings, the same reference numerals represent the same or similar components, or the same or similar steps.

The objects and functions of the present invention and methods for achieving the objects and functions will be clarified by referring to the exemplary embodiments. However, the present invention is not limited to the exemplary embodiments disclosed below; it can be implemented in various forms. The essence of the description is only to help those skilled in the relevant art comprehensively understand the specific details of the present invention.

The present invention is described in detail in conjunction with schematic diagrams. When describing the embodiments of the present invention in detail, for the convenience of explanation, the cross-sectional view showing the structure of the device will not be partially enlarged according to the general scale, and the schematic diagram is only an example, which should not limit this invention. scope of invention protection. In addition, the three-dimensional space dimensions of length, width and depth should be included in actual production.

The present invention provides a device for adjusting the optical path of a flow cytometer without a liquid path. Fig. 1 is a schematic diagram of a system block diagram of the device for adjusting the optical path of the flow cytometer without a liquid path. As shown in Fig. 1, the device Including: irradiation light source 101, irradiation spot shaping optical path 102, standard microsphere rotating device 103, non-forward scattered light beam shaping optical path 104, non-forward scattered light detection circuit 105, forward scattered light and fluorescent light beam shaping optical path 106, front Scattered light detection circuit 107, multicolor fluorescence spectroscopic optical path 108 and multi-channel fluorescence detection circuit 109, wherein,

The illumination light source 101 provides an excitation light source for fluorescence excitation;

The illumination spot shaping optical path 102 is used to compress the light source beam into an illumination spot of a certain size;

The standard microsphere rotating device 103 is used to rotate the disc loaded with standard microspheres so as to simulate single cells passing through the irradiation spot one by one;

The non-forward scattered light beam shaping optical path 104 is used to focus the non-forward scattered light beam within a certain range;

The non-forward scattered light detection circuit 105 is used for performing photoelectric conversion on the non-forward scattered light and realizing parameter extraction for the generated electric pulse signal;

The forward scattered light and fluorescent beam shaping optical path 106 is used to focus the forward scattered light and fluorescent beam within a certain range;

The forward scattered light detection circuit 107 is used to photoelectrically convert the forward scattered light and extract parameters from the generated electric pulse signal;

The multicolor fluorescence spectroscopic optical path 108 is used to separate the fluorescence signals in each wavelength range and transmit them to corresponding detection circuits;

The multi-channel fluorescence detection circuit 109 is used for photoelectric conversion of the fluorescence signals of each channel and parameter extraction of the generated electrical pulse signals.

Fig. 2 is a schematic diagram of a preferred embodiment of an optical path adjustment device for a flow cytometer of the present invention without a liquid path. It schematically shows the illumination light source 101 , the standard microsphere rotating device 103 , the non-forward scattered light beam shaping optical path 104 and the forward scattered light and fluorescent light beam shaping optical path 106 .

Preferably, a laser with a certain power and wavelength is used as the irradiation light source 101, and the laser emission beam is compressed and shaped through the irradiation spot shaping optical path 102. The irradiation spot shaping optical path 102 realizes two The size of the spot in two orthogonal directions is compressed, and at the same time, the moving microsphere is irradiated at the focal point of the spot through the irradiation spot shaping optical path 102. 3a-3c are schematic diagrams of an exemplary structure of the illumination light source and the illumination spot shaping optical path shown in FIG. 2 . 3a shows the shape change of the light spot after the laser light emitted by the irradiation light source 101 passes through the shaping lens. 3b-3c are schematic diagrams of light intensity distribution of the illuminated spot after shaping shown in FIG. 3a.

Preferably, Figures 4a-4b show a schematic diagram of an implementation of the standard microsphere rotating device 103 and standard microspheres. This standard microsphere rotation device 103 comprises a turntable that is loaded with standard microsphere, a turntable driving device and a motor motion control circuit, this turntable that is loaded with standard microsphere is connected with motor motion control circuit by turntable drive device, by adjusting motor The rotation speed controls the rotation speed of the turntable, thereby changing the time when the microspheres are irradiated by the light spot and the time interval between adjacent microspheres being irradiated.

Preferably, the turntable loaded with standard microspheres consists of a bottom layer and a top layer, wherein there are pits for placing standard microspheres in one layer, and the standard microspheres are sealed in the pits by bonding the top layer and the bottom layer to prevent microspheres. The ball falls off during the spin. During the rotation of the turntable loaded with standard microspheres, the standard microspheres pass through the irradiated spot area one by one to form side scattered light, forward scattered light and fluorescent signals of various colors.

FIG. 5 shows a structural diagram of a specific optical path embodiment of the schematic non-forward scattered light beam shaping optical path 104 . Preferably, the non-forward scattered light beam shaping optical path is placed in the non-forward scattered angle detection area, and the non-forward scattered light within a certain angle range is focused and shaped. As shown in Figure 5, the non-forward scattered light is collected through the collection lens. The scattered light is then filtered by the blocking filter, and then passes through the focusing lens, and the shaped light beam is transmitted to the non-forward scattered light detection circuit 105 .

The non-forward scattered light detection circuit 105 realizes the photoelectric conversion of the non-forward scattered light signal and the conditioning processing of the electric pulse signal, and realizes the extraction of electric pulse parameters characterizing the characteristics of the microspheres. FIG. 6 shows a specific circuit block diagram of the non-forward scattered light detection circuit 105 . As shown in Figure 6, the non-forward scattered light detection circuit 105 includes a sensor, which is used to convert the collected light signal into an electrical signal, amplify and filter the electrical pulse signal output by the sensor, and convert the conditioned electrical pulse The signal is transmitted to the subsequent analog-to-digital conversion module to be converted into a digital signal, and then input to the electric pulse parameter extraction module for extracting the required electric signal parameters.

Preferably, the forward scattered light and fluorescent beam shaping optical path 106 is placed in the direction of the irradiating light beam, focuses and shapes the forward scattered light and multi-channel fluorescent beam within a certain angle range, and transmits it to the forward scattered light detection circuit 107 . The forward scattered light detection circuit 107 realizes the photoelectric conversion of the forward scattered light signal and the conditioning processing of the electric pulse signal, and realizes the extraction of the electric pulse parameters characterizing the characteristics of the microspheres.

FIG. 7 shows a specific circuit block diagram of the forward scattered light detection circuit 107 . As shown in Figure 7, the forward scattered light detection circuit 107 includes a sensor, which is used to convert the collected optical signal into an electrical signal, amplify and filter the electrical pulse signal output by the sensor, and convert the conditioned electrical pulse signal It is transmitted to the subsequent analog-to-digital conversion module to be converted into a digital signal, and then input to the electric pulse parameter extraction module for extracting the required electric signal parameters.

FIG. 8 and FIG. 9 respectively show the structure diagram of a specific embodiment of the optical path of the schematic multicolor fluorescence beam splitting optical path 108 . As shown in Figure 8 and Figure 9, the forward scattered light is collected by the collection lens, then split through a plurality of dichroic beam splitters, filtered by a band-pass filter, and passed through a plurality of photomultiplier tubes (PMT1- 6) Perform signal multiplication collection. Preferably, the multi-color fluorescence beam splitting optical path 108 separates the multi-channel fluorescence signals through a series of optical devices, and transmits the separated fluorescent light beams of various colors to the corresponding multi-channel fluorescence detection circuit 109 .

The multi-channel fluorescence detection circuit 109 realizes the photoelectric conversion of the fluorescence signal and the conditioning processing of the electric pulse signal, and realizes the extraction of the electric pulse parameters characterizing the characteristics of the microspheres. FIG. 10 shows a specific circuit block diagram of the multi-channel fluorescence detection circuit 109 . As shown in Figure 10, the multi-channel fluorescence detection circuit 109 includes multiple fluorescence detection circuits, each of which includes a sensor for converting the collected optical signal into an electrical signal, amplifying and filtering the electrical pulse signal output by the sensor, And the conditioned electrical pulse signal is transmitted to a subsequent analog-to-digital conversion module for conversion into a digital signal, and then input to the electrical pulse parameter extraction module for extracting required electrical signal parameters.

Another aspect of the present invention provides a method for adjusting the optical path of a flow cytometer without a liquid path, comprising the steps of:

The irradiation spot shaping optical path 102 compresses the light beam generated by the irradiation light source 101 to obtain an irradiation spot of a certain size, and adjusts the focal length of the spot so that the microsphere passes through the focal point;

The standard microsphere rotating device adjusts the movement speed of the microsphere, changes the detection frequency of the microsphere and the irradiation time of the microsphere, thereby changing the frequency and duration of the corresponding light pulse and electric pulse;

The non-forward scattered light beam shaping optical path performs shaping and other processing on the non-forward scattered light, and adjusts the focal length of the optical path so that the detector in the non-forward scattered light detection circuit is at the focal point;

The non-forward scattered light detection circuit realizes the photoelectric conversion, electric pulse conditioning, analog/digital conversion and parameter extraction of non-forward scattered light, and identifies the starting point and ending point of the electric pulse;

The forward scattered light and fluorescent beam shaping optical path performs shaping and other processing on the forward scattered light and fluorescent beam, and transmits the focused beam to the multicolor fluorescent beam splitting optical path;

The multi-color fluorescence spectroscopic optical path splits the forward scattered light and the fluorescent beams of each channel according to the wavelength information, and transmits the split beams to the forward scattered light detection circuit and the corresponding multi-channel fluorescence detection circuit;

The forward scattered light detection circuit realizes the photoelectric conversion, electric pulse conditioning, analog/digital conversion and parameter extraction of forward scattered light, and identifies the starting point and ending point of the electric pulse;

The multi-channel fluorescence detection circuit realizes the photoelectric conversion, electric pulse conditioning, analog/digital conversion and parameter extraction of each fluorescence, and identifies the starting point and ending point of the electric pulse.

Other embodiments of the invention will be apparent to and understood by those skilled in the art from consideration of the specification and practice of the invention disclosed herein. The description and examples are considered exemplary only, with the true scope and spirit of the invention defined by the claims.

Claims (8)

1. a kind of optical path adjustment device of flow cytometer without fluid path situation, which is characterized in that described device includes：Irradiation light Source, irradiation spot shaping light path, standard microballoon rotating device, non-forward scattering light beam shaping light path, the inspection of non-forward scattering light Slowdown monitoring circuit, forward scattering light and fluorescent light beam shaping light path, forward scattering optical detection circuit, multicolor fluorescence light splitting optical path and mostly logical Road fluoroscopic examination circuit, wherein

The radiation source provides excitation light source for fluorescence excitation；

The irradiation spot shaping light path is used for the illumination spot of light beam of light source boil down to certain size；

The standard microballoon rotating device for make the disk for being mounted with standard microballoon rotate to simulate individual cells by One passes through irradiation light spot；

The non-forward scattering light beam shaping light path is for being focused a certain range of non-forward scattering light light beam；

The non-forward scattering optical detection circuit is used to carry out opto-electronic conversion to non-forward scattering light and believe the electric pulse of generation Number realize parameter extraction；

The forward scattering light and fluorescent light beam shaping light path be used for a certain range of forward scattering light and fluorescent light beam into Line focusing；

The forward scattering optical detection circuit is used to carry out opto-electronic conversion to forward scattering light and to the electric impulse signal of generation reality Existing parameter extraction；

The multicolor fluorescence light splitting optical path by the fluorescence signal of each wave-length coverage for being detached and being transferred to corresponding detection Circuit, the multichannel fluoroscopic examination circuit are used to carry out opto-electronic conversion to each channel fluorescence signal and to the electric impulse signal of generation Realize parameter extraction.

2. optical path adjustment device according to claim 1, it is characterised in that：Made using the laser of certain power and wavelength For radiation source, and light beam is emitted to laser by irradiating spot shaping light path and carries out compression shaping, the irradiation spot shaping Light path realizes that the spot size of two orthogonal directions is compressed according to the microballoon direction of motion in standard microballoon rotating device, passes through simultaneously Adjustment spot shaping light path makes movement microballoon be irradiated at light spot focus.

3. optical path adjustment device according to claim 2, it is characterised in that：The standard microballoon rotating device includes a dress It is loaded with the turntable, a rotating blade drive and a motor motion control circuit of standard microballoon, this is mounted with standard microballoon Turntable is connected by rotating blade drive with motor motion control circuit, and the control to rotary speed is realized by adjusting motor speed System, to change microballoon by the illuminated time interval of hot spot irradiation time and adjacent microballoon.

4. optical path adjustment device according to claim 3, it is characterised in that：Be mounted with the turntable of standard microballoon by bottom and Top layer two parts form, wherein be useful for the pit of placement standard microballoon in one layer, by the bonding of top layer and bottom by standard Microballoon, which is sealed in pit, prevents microballoon from falling off in rotary course；This is mounted with the turntable rotary course Plays of standard microballoon Microballoon forms side scattered light, forward scattering light and assorted fluorescence signal by irradiating spot area one by one.

5. optical path adjustment device according to claim 4, it is characterised in that：The non-forward scattering light beam shaping light path is put It is placed in non-forward-scattering angle detection zone, shaping is focused to the non-forward scattering light within the scope of certain angle, and by shaping Beam Propagation afterwards is to non-forward scattering optical detection circuit；The non-forward scattering optical detection circuit realizes non-forward-scattering signal Opto-electronic conversion and electric impulse signal conditioning processing, and realize to characterize microballoon characteristic electric pulse parameter extract.

6. optical path adjustment device according to claim 4, it is characterised in that：The forward scattering light and fluorescent light beam shaping light Road is positioned over illumination beam direction, within the scope of certain angle forward scattering light and multichannel fluorescent light beam be focused it is whole Shape, and by the beam Propagation after shaping to forward scattering optical detection circuit and multicolor fluorescence light splitting optical path；The forward scattering light is examined Slowdown monitoring circuit, which is realized, handles the opto-electronic conversion of forward-scattering signal and the conditioning of electric impulse signal, and realizes special to characterization microballoon The electric pulse parameter of property extracts.

7. optical path adjustment device according to claim 6, it is characterised in that：The multicolor fluorescence light splitting optical path passes through a series of Optical devices detach multichannel fluorescence signal, and it is glimmering that the assorted fluorescent light beam after separation is transferred to corresponding multichannel Optical detection circuit；The multichannel fluoroscopic examination circuit is realized to the opto-electronic conversion of fluorescence signal and the conditioning of electric impulse signal Reason, and realize and the electric pulse parameter for characterizing microballoon characteristic is extracted.

8. a kind of optical path adjusting method of flow cytometer without fluid path situation, including step：

Irradiation spot shaping light path carries out compression processing to the light beam that radiation source generates, and obtains the irradiation hot spot of certain size, And adjusting hot spot focal length makes microballoon pass through from focal point；

Standard microballoon rotating device adjusts the movement velocity of microballoon, changes microballoon detection frequency and microballoon irradiation time, to change Become corresponding light pulse and electrical impulse frequency and duration；

Non- forward scattering light beam shaping light path carries out Shape correction to non-forward scattering light, and adjustment light path focal length makes non-forward direction dissipate The detector penetrated in optical detection circuit is in focal position；

Non- forward scattering optical detection circuit realizes the opto-electronic conversion to non-forward scattering light, electric pulse conditioning, analog/digital conversion and ginseng Number extraction process, identifies the starting point and end point of electric pulse；

Forward scattering light and fluorescent light beam shaping light path carry out Shape correction to forward scattering light and fluorescent light beam, and will be after focusing Beam Propagation to multicolor fluorescence light splitting optical path；

Forward scattering light and each channel fluorescence light beam are carried out light-splitting processing by multicolor fluorescence light splitting optical path according to wavelength information, and will Beam Propagation after light splitting is to forward scattering optical detection circuit and corresponding multichannel fluoroscopic examination circuit；

Forward scattering optical detection circuit realizes that the opto-electronic conversion to forward scattering light, electric pulse improve, analog/digital conversion and parameter carry Processing is taken, identifies the starting point and end point of electric pulse；

Multichannel fluoroscopic examination circuit realizes that the opto-electronic conversion to each fluorescence, electric pulse improve, at analog/digital conversion and parameter extraction Reason, identifies the starting point and end point of electric pulse.