CN113063759B - Somatic cell laser-induced fluorescence detection method based on hemispherical space compound eye structure - Google Patents
Somatic cell laser-induced fluorescence detection method based on hemispherical space compound eye structure Download PDFInfo
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Abstract
The invention discloses a somatic cell laser-induced fluorescence detection method based on a hemispherical space compound eye structure, which comprises the following steps of: preparing a sample, selecting a corresponding antibody according to an antigen molecule to be measured, and performing fluorescent marking on the antibody by using a fluorescent agent to prepare a sample cell suspension; sample injection, namely adding the sample cell suspension into a spherical compound eye structure in a single cell flow mode; the method comprises the steps of compound eye immune LIF detection, wherein a plurality of small eye light path structures are uniformly arranged on the spherical surface of the spherical compound eye structure, the small eye light path structures on the spherical compound eye structure are irradiated to cells to be detected, LIF signals generated at the intersection points of the small eye optical axes of the small eye light path structures and the cells to be detected are measured, and the body distribution of antigen molecules of the cells to be detected is inverted; and 4, LIF signal processing, namely repeating the step 3 every fixed time t to obtain a series of LIF signals changing along with time, and ending detection after the preset detection time M is completed.
Description
Technical Field
The invention relates to a laser-induced fluorescence detection method, in particular to a single-cell three-dimensional molecular body distribution detection instrument method based on compound eye bionic optics.
Background
Cells are the fundamental units that constitute the structure of living organisms and that fulfill their vital functions. From bacteria, blue algae, protozoa and other multicellular organisms, which are composed of single cells, to higher animals and plants, the different types of cells with different functions and different numbers form simple to complex life units of organism tissues, organs, systems and the like, thereby realizing the diversity of life activities.
The cell volume is small, most typically between 1 and 30 microns in diameter, and individual larger cells can be as large as the order of centimeters. The internal structure of the cells can be seen by means of an optical microscope, which is required for finer structures of the organelles. Because the cells have functions of movement, nutrition, reproduction and the like, the dynamic detection of organic and inorganic molecules in the cells has important effect on understanding physiological activities of the cells, and contains information of rich vital activities.
The method of displaying and detecting antigen or hapten substances in cells or tissues by immunofluorescence technology is called immunofluorescence cell (or tissue) chemistry technology, and the technology can be used for detecting specific protein molecules in cells by combining with microscopic imaging, but the conventional method cannot obtain the space dynamic change of specific molecules to be detected in cells, so that the requirement of dynamic detection of specific molecules in cells cannot be met, such as the transportation or action process of the specific molecules in cells; the process by which viral molecules bind to receptors inside cells and erode them; the course of action of drug molecules within cells, and the like.
For example, in the published prior patent application "an immunofluorescence detection device" (chinese patent publication No. CN 201917571U), an immunofluorescence detection device is disclosed, which structurally comprises a laser light source, an excitation light path, a moving platform, a fluorescence light path, a photoelectric conversion system, a control system, a data processing system, and a reagent card, wherein the laser light source, the excitation light path, and the fluorescence light path are assembled on the same optical base, the optical base and the moving platform are mounted on the device base, and the laser light source is a semiconductor laser tube fixed in an inner sleeve and an outer sleeve.
The defects of the existing immunofluorescence detection technical scheme are very remarkable, the detection of cells from a single fixed light path angle can only be realized, the detection result is planar and non-real-time, and the spatial dynamic change of specific molecules to be detected in the cells can not be realized, so that the requirements for the dynamic detection of the specific molecules in the cells can not be met.
Disclosure of Invention
The invention aims to overcome the defects of the existing immunofluorescence detection technical scheme, and provides a somatic cell laser-induced fluorescence detection method based on a hemispherical space compound eye structure.
In order to achieve the above purpose, the present invention adopts the following technical scheme:
the invention discloses a somatic cell laser-induced fluorescence detection method based on a hemispherical space compound eye structure, which comprises the following steps of:
LIF data corresponding to a certain moment are obtained by analyzing the intensity of fluorescence at the intersection point of the optical axis of each small eye and the cell to be detected, and the body distribution of the antigen molecules of the cell to be detected at the moment is obtained after calculating all small eyes on the spherical compound eye structure; the same analysis method is adopted for M moments, so that the dynamic change of the body distribution of antigen molecules of the cells to be detected along with time can be obtained; the dynamic change can be visually displayed by using system software in a three-dimensional space diagram mode.
Preferably, the step 1 further includes:
selecting a corresponding antibody according to an antigen molecule to be measured, and performing fluorescent marking on the selected antibody by using a fluorescent agent; the marked fluorescent antibody is directly added on a certain amount of cell samples, and after dyeing at a certain temperature and for a certain time, the unreacted excessive fluorescent antibody is washed off by water, so that the cell suspension of the samples is prepared.
Preferably, the step 2 further includes:
starting a flowing liquid by a flow type sample injector to form a sheath flow; the sample cell suspension is sprayed out of the capillary tube and flows together with the sheath flow, single cell flow is formed in the middle of the flow tube, and the flowing cell to be tested is electrified by the electrifying head at the outlet of the flow tube and is provided with uniform negative charge;
the spherical compound eye structure comprises an upper hemispherical compound eye cover body and a lower hemispherical compound eye cover body, symmetrical positive fields are arranged in the spherical compound eye structure, after entering the inner side of the spherical compound eye structure, the main controller sends out instructions to start the translational controller, the translational controller controls the lower hemispherical compound eye to move upwards until the upper hemispherical compound eye cover body and the lower hemispherical compound eye cover body are spliced to form the spherical compound eye structure, and cells to be detected move until the mass centers of the cells are approximately coincident with the sphere centers, and detection is started at the moment.
Preferably, the step 3 further includes:
the main controller starts the laser and the array driving circuit, laser emitted by the laser is transmitted along an input optical axis, collimated and expanded by the collimating lens, each small lens is focused into N input optical fibers, enters the small-eye optical fibers in the small-eye optical path structure through the Y-shaped connector, and is converged to cells to be detected at the sphere center by the small-eye lens group;
the number of antigen molecules at the intersection point of each small eye optical axis and the cell to be detected is different, and fluorescent-labeled antibody molecules linked on the antigen molecules generate fluorescence under the excitation of laser, and the intensity of the fluorescence reflects the number of the antigen molecules; the body distribution of antigen molecules of the cells to be detected can be inverted by measuring LIF signals generated at the intersection point of the optical axis of the small eye and the cells to be detected;
under the excitation of laser, LIF signals generated at antigen molecules are collected and focused to the end face by the signals of each small glasses group, then enter small-eye optical fibers, enter output optical fibers through Y-shaped connectors, the output optical fibers send the signals to a photoelectric tube array, and a photoelectric diode converts fluorescent signals into electric signals; the array driving circuit amplifies and analog-to-digital converts the electric signals output by the photoelectric tube array and sends the electric signals to the main controller for storage and analysis.
According to the technical scheme, the cell detection instrument based on the bionic compound eye is provided. The spherical compound eye structure formed by splicing the upper hemisphere and the lower hemisphere and the electrodes on the inner surface form a uniform electric field in the sphere, when the electric field force is balanced, uniformly charged cells can be fixed at the position of the sphere center, and the body distribution of the molecules of interest in the cells can be conveniently detected. Each small eye adopts an LIF light path formed by a small eye light path structure, an excitation light path structure and a fluorescence light path structure to perform laser excitation and immunofluorescence reception, and the dynamic detection of the spatial distribution and the change of specific antigen molecules in cells can be realized, so that the research requirement on specific physiological movement of the cells is met. Meanwhile, the small eye light path structure is arranged on the spherical compound eye structure, and dynamic detection of cells to be detected on all angles in a three-dimensional space can be realized. The dynamic change of the body distribution of the antigen molecules of the cells to be detected along with the time can be obtained by detecting at fixed time intervals; the dynamic change can be visually displayed by using system software in a three-dimensional space diagram mode.
Drawings
FIG. 1 is a general structural diagram of a hemispherical space compound eye somatic cell laser-induced fluorescence detector of the invention.
Fig. 2 is a diagram showing a structure of a microlens array according to the present invention.
Fig. 3 is a diagram showing a structure of a photoelectric cell array according to the present invention.
Fig. 4 is a diagram of the structure of the upper hemisphere compound eye mask of the present invention.
Fig. 5 is a block diagram of a small eye optical path structure in the present invention.
FIG. 6 is a graph showing the relationship between the cells to be tested and the optical axis of the eyes in the present invention.
Fig. 7 is a side view of the upper hemispherical complex eye shield of the present invention.
Fig. 8 is a view showing a meridian plane structure of the upper hemisphere compound eye mask body in the present invention.
The drawing is marked: 1. a lower hemisphere compound eye cover body; 2. a collection plate; 3. a small eye optical fiber; 4. a Y-type joint; 5. an output optical fiber; 6. a photocell array; 7. an array driving circuit; 8. a microlens array; 9. a collimator lens; 10. a laser; 11. an input optical axis; 12. a flow injector; 13. a sample port; 14. sheath flow; 15. a main controller; 16. a single cell stream; 17. a test cell; 18. powering on the electric head; 19. an input optical fiber; 20. a lenslet; 21. a band-pass filter; 22. a photodiode; 23. installing small holes; 24. an optical axis of the small eye; 25. anterior ocular lens; 26. a retrobulbar lens; 27. an end face; 28. an antigen molecule; 29. a pole; 30. an included angle between the surfaces; 31. meridian plane; 32. a center of sphere; 33. an optical axis included angle; 34. an upper hemisphere compound eye cover body; 35. a translational controller; 36. an inner surface electrode; 37. a capillary tube; 38. and a flow tube.
Detailed Description
The invention is further described below with reference to the drawings and detailed description.
As shown in fig. 1, the somatic cell laser-induced fluorescence detection method based on the hemispherical space compound eye structure is realized based on the hemispherical space compound eye somatic cell LIF detector provided by the embodiment of the application. The hemispherical space compound eye like somatic cell LIF detector comprises an excitation light path structure, a fluorescence light path structure, a flow sample injector 12 and a spherical compound eye structure. The spherical compound eye structure comprises an upper hemispherical compound eye cover body 1 and a lower hemispherical compound eye cover body 34 which can be closely spliced. The lower hemispherical compound eye cover 34 is provided with a translational controller 35, and the translational controller 35 controls the lower hemispherical compound eye cover 34 to move up and down to be close to or far away from the upper hemispherical compound eye cover 1.
As shown in fig. 1 and 2, the excitation light path structure includes a laser 10, a collimator lens 9, a microlens array 8 and an input optical fiber 19, which are sequentially connected, the microlens array 8 includes a plurality of lenslets 20 arranged in an array, and the end of each input optical fiber 19 is connected with a single lenslet 20 in a butt joint manner.
As shown in fig. 1 and 3, the fluorescent light path structure includes an output optical fiber 5, a photocell array 6 and an array driving circuit 7, the photocell array 6 includes a bandpass filter 21 and a photodiode 22, and the end of the output optical fiber 5 is connected with a single group of bandpass filter 21 and photodiode 22.
As shown in fig. 4, 5 and 6, the upper hemispherical complex eye shield body 1 and the lower hemispherical complex eye shield body 34 are arranged with a small eye light path structure. The small eye light path structures are evenly distributed on all meridian planes 31 of the spherical compound eye structure, the small eye light axes 24 of all the small eye light path structures point to the sphere centers 32 of the spherical compound eye structure, the light axis included angles 33 between all the adjacent small eye light axes 24 are equal, and the plane included angles 30 between all the adjacent meridian planes 31 are equal.
As shown in fig. 5, the optical path structure for the small eye comprises an optical fiber for the small eye 3, a front lens for the small eye 25 and a rear lens for the small eye 26, wherein the upper hemispherical compound eye cover body 1 and the lower hemispherical compound eye cover body 34 are respectively provided with a small mounting hole 23, the optical fiber for the small eye 3 is inserted into the small mounting hole 23 along the optical axis for the small eye 24, the front lens for the small eye 25 is arranged on the end face of the inner end of the small mounting hole 23, and the rear lens for the small eye 26 is arranged in the middle of the small mounting hole 23.
The output optical fiber 5 and the input optical fiber 19 are connected to the eyelet optical fiber 3 through the Y-joint 4.
N input optical fibers 19 are arranged according to the sequence of the small eye numbers and are arranged on the micro lens array 8 to be matched with the small lenses 20 with the same number as N; the microlens array 8 is perpendicular to the input optical axis 11 and centered on the input optical axis 11; the laser 10 is used for emitting pump laser light for exciting LIF fluorescence signals; the collimating lens 9 collimates the laser beam output by the laser 10 and covers the micro lens array 8; each lenslet 20 focuses the collimated laser light into a respective input optical fiber 19; n output optical fibers 5 are arranged according to the sequence of the small eye numbers and are arranged on the photoelectric tube array 6 to be matched with the photoelectric diodes 22 with the same number as N; each photodiode 22 covers a bandpass filter 21, the passband wavelength of which corresponds to the fluorescence band generated by the pump laser exciting the fluorescence labeled antibody, the photodiodes 22 converting the fluorescence signal into an electrical signal; the array driving circuit 7 is used for driving the photoelectric tube array 6 to work, and amplifying and analog-digital converting the electric signal output by the photoelectric tube array 6, and then sending the electric signal to the main controller 15 for storage and analysis.
The excitation light path structure, the fluorescence light path structure and the small eye light path structure form an LIF light path, so that each small eye on the spherical compound eye structure can perform laser excitation and immunofluorescence reception, and dynamic detection on the spatial distribution and the change of specific antigen molecules in cells can be realized, thereby meeting the research requirements on specific physiological movements of the cells. Meanwhile, the small eye light path structure is arranged on the spherical compound eye structure, and dynamic detection of cells to be detected on all angles in a three-dimensional space can be realized.
The main controller 15 is used for sending control instructions to the laser 10, the translation controller 35, the flow type injector 12, the power-on head 18, the laser 10, the array driving circuit 7 and the collecting plate 2, controlling the work of the control instructions and also is used for receiving data of the array driving circuit 7 for storage and analysis.
As shown in fig. 1, 7 and 8, the upper hemispherical complex eye shield body 1 and the lower hemispherical complex eye shield body 34 are provided with two poles 29, and the inner surfaces are formed by metal plating, so as to form inner surface electrodes 36. When the upper hemispherical compound eye cover body 1 and the lower hemispherical compound eye cover body 34 are closed and spliced, the inner surface electrode 36 is positively charged, so that a symmetrical inner electric field is formed in the spherical compound eye structure. The internal electric field is positive. The outlet of the flow injector 12 is provided with an electrical head 18 for negatively charging the cells to be measured. The flow sampler 12 comprises a capillary tube 37 and a flow tube 38, and a collecting plate 2 is arranged on the other side of the spherical compound eye structure, and the collecting plate 2 is positively charged.
When in sample injection, the flow type sample injector starts flowing liquid to flow along the periphery in the flow pipe to form sheath flow; the sample cell suspension is ejected from the capillary 37 and flows together with the sheath flow 14 to form a single cell flow 16 in the middle of the flow tube, and the outgoing cells 17 to be tested are electrified by the electrifying head at the outlet of the flow tube 38 and are uniformly negatively charged. After entering the interior of the spherical compound eye structure, the upper hemispherical compound eye cover body and the lower hemispherical compound eye cover body are spliced, under the action of an internal spherical symmetrical electric field in the compound eye, the cells to be detected with uniform negative charges move until the mass centers of the cells to be detected are approximately coincident with the sphere centers, and the cells are approximately in an equilibrium state under the action of electrostatic force, so that detection can be started; after detection, separating the lower hemisphere compound eye from the upper hemisphere compound eye, and approaching the cells to be detected to the collecting plate under the action of the positively charged collecting plate until the cells are attached to the upper collecting plate.
The invention discloses a somatic cell laser-induced fluorescence detection method based on a hemispherical space compound eye structure, which comprises the following steps of:
Specifically, the step 1 further includes:
selecting a corresponding antibody according to an antigen molecule (28) to be measured, and performing fluorescent marking on the selected antibody by using a fluorescent agent; the marked fluorescent antibody is directly added on a certain amount of cell samples, and after dyeing at a certain temperature and for a certain time, the unreacted excessive fluorescent antibody is washed off by water, so that the cell suspension of the samples is prepared.
And 2, sample injection, namely adding the sample cell suspension into a spherical compound eye structure in a single cell flow (16) mode, enabling the cells (17) to be detected to have uniform negative charge through upper nodding, wherein symmetrical positive fields are arranged in the spherical compound eye structure, and detecting after the centers of mass of the cells (17) to be detected are approximately coincident with the centers of spheres.
Specifically, the step 2 further includes:
the flow injector (12) starts flowing liquid to form sheath flow (14); the sample cell suspension is sprayed out of the capillary tube (37) and flows together with the sheath flow (14), a single cell flow (16) is formed in the middle of the flow tube (38), and the flowing cell (17) to be tested is electrified by the electrifying head (18) at the outlet of the flow tube (38) and is provided with uniform negative charges.
The spherical compound eye structure comprises an upper hemispherical compound eye cover body (34) and a lower hemispherical compound eye cover body (1), symmetrical positive fields are arranged in the spherical compound eye structure, after entering the inner side of the spherical compound eye structure, a main controller (15) sends out instructions to start a translational controller (35), the translational controller (35) controls the lower hemispherical compound eye to move upwards until the upper hemispherical compound eye cover body (34) and the lower hemispherical compound eye cover body (1) are spliced to form the spherical compound eye structure, and a cell (17) to be detected moves until the centroid of the cell is approximately coincident with the sphere center, and detection is started at the moment.
Specifically, the step 3 further includes:
the main controller (15) starts the laser (10) and the array driving circuit (7), laser emitted by the laser (10) is transmitted along the input optical axis (11), collimated and expanded by the collimating mirror (9), each small lens (20) is focused into N input optical fibers (19), enters the small eye optical fibers (3) in the small eye optical path structure through the Y-shaped connector (4), and is converged to the cell (17) to be detected at the sphere center through the small eye group;
the number of antigen molecules (28) at the intersection point of each small-eye optical axis (24) and the cell (17) to be detected is different, and fluorescent-labeled antibody molecules linked on the antigen molecules (28) generate fluorescence under the excitation of laser, and the intensity of the fluorescence reflects the number of the antigen molecules (28); inverting the body distribution of the antigen molecules (28) of the test cell (17) by measuring LIF signals generated at the intersection of the small eye optical axis (24) and the test cell (17);
under the excitation of laser, LIF signals generated at the antigen molecules (28) are collected and focused to the end face by the signals of each small glasses group, then enter the small eye optical fiber (3), enter the output optical fiber (5) through the Y-shaped connector (4), the output optical fiber (5) sends the signals to the photoelectric cell array (6), and the photoelectric diode (22) converts the fluorescent signals into electric signals; the array driving circuit (7) amplifies and analog-digital converts the electric signal output by the photoelectric tube array (6) and sends the electric signal to the main controller (15) for storage and analysis.
LIF data corresponding to a certain moment are analyzed to obtain the relative number of antigen molecules (28) of the point by analyzing the fluorescence intensity at the intersection point of the optical axis (24) of each small eye and the cell (17) to be detected, and after all small eyes on the spherical compound eye structure are calculated, the body distribution of the antigen molecules (28) of the cell (17) to be detected at the moment is obtained; the same analysis method is adopted for M moments, so that the dynamic change of the body distribution of antigen molecules (28) of the cells (17) to be detected along with the time can be obtained; the dynamic change can be visually displayed by using system software in a three-dimensional space diagram mode.
According to the technical scheme, the cell detection instrument based on the bionic compound eye is provided. The spherical compound eye structure formed by splicing the upper hemisphere and the lower hemisphere and the electrodes on the inner surface form a uniform electric field in the sphere, when the electric field force is balanced, uniformly charged cells can be fixed at the position of the sphere center, and the body distribution of the molecules of interest in the cells can be conveniently detected. Each small eye adopts an LIF light path formed by a small eye light path structure, an excitation light path structure and a fluorescence light path structure to perform laser excitation and immunofluorescence reception, and the dynamic detection of the spatial distribution and the change of specific antigen molecules in cells can be realized, so that the research requirement on specific physiological movement of the cells is met. Meanwhile, the small eye light path structure is arranged on the spherical compound eye structure, and dynamic detection of cells to be detected on all angles in a three-dimensional space can be realized. The dynamic change of the body distribution of the antigen molecules of the cells to be detected along with the time can be obtained by detecting at fixed time intervals; the dynamic change can be visually displayed by using system software in a three-dimensional space diagram mode.
Claims (4)
1. A somatic cell laser-induced fluorescence detection method based on a hemispherical space compound eye structure is characterized by comprising the following steps:
step 1, preparing a sample, namely selecting a corresponding antibody according to an antigen molecule (28) to be measured, and performing fluorescent marking on the antibody by using a fluorescent agent to prepare a sample cell suspension;
step 2, sample injection, namely adding sample cell suspension into a spherical compound eye structure in a single cell flow (16) mode, enabling cells (17) to be detected to have uniform negative charge through an electrifying head, wherein symmetrical positive fields are arranged in the spherical compound eye structure, and detecting after the centers of mass of the cells (17) to be detected are approximately coincident with the centers of spheres;
step 3, compound eye immune LIF detection, wherein a plurality of small eye light path structures are uniformly arranged on the spherical surface of the spherical compound eye structure, and the small eye light path structures are connected with an excitation light path structure and a fluorescence light path structure; the method comprises the steps that laser output by an excitation light path structure irradiates a cell (17) to be detected through a small eye light path structure on a spherical compound eye structure, LIF signals generated at the intersection point of a small eye light axis (24) of the small eye light path structure and the cell (17) to be detected are measured through a fluorescence light path structure to invert the body distribution of antigen molecules (28) of the cell (17) to be detected, and the excitation light path structure, the fluorescence light path structure and the small eye light path structure form a LIF light path, so that each small eye on the spherical compound eye structure can perform laser excitation and immunofluorescence reception, and dynamic detection of the spatial distribution and the change of specific antigen molecules in the cell can be realized;
step 4, LIF signal processing, namely repeating the step 3 every other fixed time t to obtain a series of LIF signals changing along with time, and ending detection after the preset detection time M is completed;
LIF data corresponding to a certain moment are analyzed to obtain the relative number of antigen molecules (28) of the point by analyzing the fluorescence intensity at the intersection point of the optical axis (24) of each small eye and the cell (17) to be detected, and after all small eyes on the spherical compound eye structure are calculated, the body distribution of the antigen molecules (28) of the cell (17) to be detected at the moment is obtained; the same analysis method is adopted for all M moments, so that the dynamic change of the body distribution of antigen molecules (28) of the cells (17) to be detected along with the time can be obtained.
2. The method for detecting somatic cell laser-induced fluorescence based on hemispherical spatial compound eye structure according to claim 1, wherein the step 1 further comprises:
selecting a corresponding antibody according to an antigen molecule (28) to be measured, and performing fluorescent marking on the selected antibody by using a fluorescent agent; the marked fluorescent antibody is directly added on a certain amount of cell samples, and after dyeing at a certain temperature and for a certain time, the unreacted excessive fluorescent antibody is washed off by water, so that the cell suspension of the samples is prepared.
3. The method for detecting somatic cell laser-induced fluorescence based on hemispherical spatial compound eye structure according to claim 1, wherein the step 2 further comprises:
the flow injector (12) starts flowing liquid to form sheath flow (14); the sample cell suspension is sprayed out of a capillary tube (37) and flows together with a sheath flow (14), a single cell flow (16) is formed in the middle of a flow tube (38), and the flowing cell (17) to be tested is electrified by an electrifying head (18) at the outlet of the flow tube (38) and is provided with uniform negative charge;
the spherical compound eye structure comprises an upper hemispherical compound eye cover body (34) and a lower hemispherical compound eye cover body (1), symmetrical positive fields are arranged in the spherical compound eye structure, after entering the inner side of the spherical compound eye structure, a main controller (15) sends out instructions to start a translational controller (35), the translational controller (35) controls the lower hemispherical compound eye to move upwards until the upper hemispherical compound eye cover body (34) and the lower hemispherical compound eye cover body (1) are spliced to form the spherical compound eye structure, and a cell (17) to be detected moves until the centroid of the cell is approximately coincident with the sphere center, and detection is started at the moment.
4. The method for detecting somatic cell laser-induced fluorescence based on hemispherical spatial compound eye structure according to claim 1, wherein the step 3 further comprises:
the main controller (15) starts the laser (10) and the array driving circuit (7), laser emitted by the laser (10) is transmitted along the input optical axis (11), collimated and expanded by the collimating mirror (9), each small lens (20) is focused into N input optical fibers (19), enters the small eye optical fibers (3) in the small eye optical path structure through the Y-shaped connector (4), and is converged to the cell (17) to be detected at the sphere center through the small eye group;
the number of antigen molecules (28) at the intersection point of each small-eye optical axis (24) and the cell (17) to be detected is different, and fluorescent-labeled antibody molecules linked on the antigen molecules (28) generate fluorescence under the excitation of laser, and the intensity of the fluorescence reflects the number of the antigen molecules (28); inverting the body distribution of the antigen molecules (28) of the test cell (17) by measuring LIF signals generated at the intersection of the small eye optical axis (24) and the test cell (17);
under the excitation of laser, LIF signals generated at the antigen molecules (28) are collected and focused to the end face by the signals of each small glasses group, then enter the small eye optical fiber (3), enter the output optical fiber (5) through the Y-shaped connector (4), the output optical fiber (5) sends the signals to the photoelectric cell array (6), and the photoelectric diode (22) converts the fluorescent signals into electric signals; the array driving circuit (7) amplifies and analog-digital converts the electric signal output by the photoelectric tube array (6) and sends the electric signal to the main controller (15) for storage and analysis.
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