CN113063759A - 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: preparing a sample, namely selecting a corresponding antibody according to an antigen molecule to be measured, and carrying out fluorescence labeling on the antibody by using a fluorescent agent to prepare a sample cell suspension; sample introduction, namely adding a sample cell suspension into a spherical compound eye structure in a single cell flow mode; compound eye immune LIF detection, wherein a plurality of small eye light path structures are uniformly distributed on the spherical surface of the spherical compound eye structure, the spherical compound eye structure irradiates to a cell to be detected through the small eye light path structures on the spherical compound eye structure, and LIF signals generated at the intersection point of a small eye light axis of the small eye light path structure and the cell to be detected are determined to be inverted to perform the volume distribution of antigen molecules of the cell to be detected; and 4, LIF signal processing, repeating the step 3 every other fixed time t to obtain a series of LIF signals changing along with time, and finishing detection after the preset detection time number M is finished.
Description
Technical Field
The invention relates to a laser-induced fluorescence detection method, in particular to a unicellular three-dimensional molecular body distribution detection instrument method based on compound eye bionic optics.
Background
Cells are the basic units that make up the structure of a living body and perform vital functions. From bacteria, blue algae, protozoa, and multi-cellular organisms such as higher animals and plants, which are composed of single cells, different types of cells with different functions and different numbers constitute simple to complex life units such as tissues, organs, systems, and the like of organisms, thereby realizing diversity of life activities.
The cells are small in volume, with most typical cells being 1 to 30 microns in diameter, and individual larger cells can be as large as a centimeter. The internal structure of the cell can be seen with the aid of an optical microscope, which is required for the more subtle structure of the organelles. Because the cells have the functions of movement, nutrition, reproduction and the like and contain rich information of life activities, the dynamic detection of organic and inorganic molecules in the cells plays an important role in understanding the physiological activities of the cells.
The method of displaying and checking antigen or hapten substances in cells or tissues by using an immunofluorescence technique is called an immunofluorescence cell (or tissue) chemical technique, and the specific protein molecules in the cells can be detected by combining the technique with microscopic imaging, but the conventional method cannot obtain the space dynamic change of the specific molecules to be detected in the cells, so that the requirement on the dynamic detection of the specific molecules in the cells cannot be met, such as the transportation or action process of the intracellular protein molecules on the certain molecules; the process by which viral molecules bind to receptors inside cells and erode cells; the action process of the drug molecules in the cell, etc.
For example, in the application of the published prior patent application "an immunofluorescence detection apparatus" (chinese patent publication No. CN 201917571U), an immunofluorescence detection apparatus is disclosed, which comprises a laser source, an excitation light path, a mobile platform, a fluorescence light path, a photoelectric conversion system, a control system, a data processing system, and a reagent card, wherein the laser source, the excitation light path, and the fluorescence light path are assembled on the same optical base, the optical base and the mobile platform are mounted on the apparatus base, and the laser 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 obvious, the cell can only be detected from the angle of a single fixed light path, the detection result is planar and non-real-time, and the dynamic change of the space of a specific molecule to be detected in the cell cannot be realized, so that the requirement for dynamic detection of the specific molecule in the cell cannot 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 purpose, the invention adopts the following technical scheme:
the invention relates to a somatic cell laser-induced fluorescence detection method based on a hemispherical space compound eye structure, which comprises the following steps of:
corresponding to LIF data at a certain moment, obtaining the relative number of antigen molecules at the point by analyzing the intensity of fluorescence at the intersection of each ommatidium optical axis and the cell to be detected, and obtaining the volume distribution of the antigen molecules of the cell to be detected at the moment after calculating all ommatidium on the spherical compound eye structure; the same analysis method is adopted for M moments, and the dynamic change of the distribution of the antigen molecules of the cells to be detected along with the time can be obtained; the dynamic change can be visually displayed by a three-dimensional space dynamic diagram by system software.
Preferably, the step 1 further comprises:
selecting a corresponding antibody according to an antigen molecule to be measured, and carrying out fluorescence labeling on the antibody by using a fluorescent agent after the antibody is selected; directly adding the marked fluorescent antibody on a certain amount of cell samples, dyeing at a certain temperature and time, and washing off the excessive fluorescent antibody which does not participate in the reaction by using water to prepare the sample cell suspension.
Preferably, the step 2 further comprises:
starting the flowing liquid by the flow type sample injector to form sheath flow; the sample cell suspension is sprayed out of the capillary tube and flows together with the sheath flow to form single cell flow in the middle of the flow tube, and the flowing out cell to be detected is electrified by the upper electrode at the outlet of the flow tube and is provided with uniform negative charges;
the spherical compound eye structure comprises an upper hemispherical compound eye cover body and a lower hemispherical compound eye cover body, wherein symmetrical positive electric fields are arranged in the spherical compound eye structure, after the positive electric fields enter the inner side of the spherical compound eye structure, a main controller sends out an instruction to start a flat controller, the flat 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, a cell to be detected moves until the mass center of the cell to be detected is approximately coincident with the sphere center, and detection is started at the moment.
Preferably, the step 3 further comprises:
the main controller starts a laser and an array driving circuit, laser emitted by the laser is transmitted along an input optical axis, collimated and expanded by a collimating lens, focused by each small lens to enter N input optical fibers, then enters the small-eye optical fibers in the small-eye optical path structure through a Y-shaped joint, and then is converged to cells to be detected at the center of the sphere through the small-eye lens group;
the number of antigen molecules at the intersection of each ommatidium optical axis and a cell to be detected is different, fluorescence is generated by the fluorescence-labeled antibody molecules linked on the antigen molecules under the excitation of laser, and the intensity of the fluorescence reflects the number of the antigen molecules; the distribution of the antigen molecules of the cells to be detected can be inverted by measuring the LIF signal generated at the intersection 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 each small-eye lens group signal, then enter small-eye optical fibers and then enter output optical fibers through Y-shaped joints, 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 converts the electric signal output by the photoelectric tube array and then sends the electric signal to the main controller for storage and analysis.
The technical scheme of the invention is based on a bionic compound eye cell detection instrument. The spherical compound eye structure formed by splicing the upper hemispherical compound eye cover body and the lower hemispherical compound eye cover body and the inner surface electrode form an in-sphere uniform electric field, and when the electric field force is balanced, cells which are uniformly charged can be fixed at the center of the sphere, so that the body distribution of the molecules of interest in the cells can be conveniently detected. Each small eye is provided with an LIF light path formed by a small eye light path structure, an excitation light path structure and a fluorescence light path structure to carry out laser excitation and immunofluorescence receiving, so that the dynamic detection of the spatial distribution and the change of specific antigen molecules in cells can be realized, and the research requirement on specific physiological movement of the cells is met. Meanwhile, the ommatidium light path structure is arranged on the spherical compound eye structure, so that dynamic detection of cells to be detected in each angle of a three-dimensional space can be realized. By detecting at fixed time intervals, the dynamic change of the distribution of the antigen molecules of the cells to be detected along with the time can be obtained; the dynamic change can be visually displayed by a three-dimensional space dynamic diagram by system software.
Drawings
Fig. 1 is a general structural diagram of the laser-induced fluorescence detector for the hemispheric space compound eye somatic cells of the present invention.
FIG. 2 is a view showing a structure of a microlens array according to the present invention.
FIG. 3 is a diagram of a photocell array of the present invention.
Fig. 4 is a structural view of an upper hemispherical compound eye mask body in the present invention.
Fig. 5 is a structural diagram of the optical path structure of the ommatidium of the present invention.
FIG. 6 is a diagram showing the relationship between the test cells and the optical axis of the ommatidium.
Fig. 7 is a side view of an upper hemispherical compound eye mask body in accordance with the present invention.
Fig. 8 is a meridian plane structure view of the upper hemispherical compound eye cover body in the present invention.
The figure is marked with: 1. a lower hemispherical compound eye mask body; 2. a collection plate; 3. a small-eye optical fiber; 4. a Y-shaped joint; 5. an output optical fiber; 6. an array of photocells; 7. an array driving circuit; 8. a microlens array; 9. a collimating mirror; 10. a laser; 11. an input optical axis; 12. a flow sample injector; 13. a sample port; 14. a sheath flow; 15. a main controller; 16. a single cell stream; 17. a cell to be tested; 18. a power-on head; 19. an input optical fiber; 20. a lenslet; 21. a band-pass color filter; 22. a photodiode; 23. mounting small holes; 24. the axis of the ommatidium; 25. a small anterior ocular lens; 26. a posterior lens of the ommatidium; 27. an end face; 28. an antigenic molecule; 29. a pole; 30. included angle between the surfaces; 31. meridian plane; 32. a spherical center; 33. an optical axis included angle; 34. an upper hemispherical compound eye mask body; 35. a translation controller; 36. an inner surface electrode; 37. a capillary tube; 38. a flow tube.
Detailed Description
The invention is further described with reference to the following figures and detailed description.
As shown in fig. 1, the laser-induced fluorescence detection method for somatic cells based on a hemispherical space compound eye structure according to the present invention is implemented based on a hemispherical space compound eye somatic cell LIF detector provided in an embodiment of the present invention. The LIF detector for the hemispheric space compound eye somatic cells comprises an excitation light path structure, a fluorescence light path structure, a flow type 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 spliced closely. The lower hemispherical compound eye mask body 34 is provided with a flat controller 35, and the flat controller 35 controls the lower hemispherical compound eye mask body 34 to move up and down to approach or be away from the upper hemispherical compound eye mask body 1.
As shown in fig. 1 and fig. 2, the excitation optical path structure includes a laser 10, a collimating mirror 9, a microlens array 8, and an input optical fiber 19, which are sequentially connected in an abutting manner, where the microlens array 8 includes a plurality of lenslets 20 arranged in an array, and a distal end of each input optical fiber 19 is connected in an abutting manner with a single lenslet 20.
As shown in fig. 1 and fig. 3, the fluorescent light path structure includes an output optical fiber 5, a photo-transistor array 6, and an array driving circuit 7, where the photo-transistor array 6 includes a band-pass filter 21 and a photodiode 22, and a single group of the band-pass filter 21 and the photodiode 22 is connected to an end of the output optical fiber 5.
As shown in fig. 4, 5 and 6, the upper hemispherical compound eye mask body 1 and the lower hemispherical compound eye mask body 34 are arranged with a small eye optical path structure. The ommatidium optical path structures are uniformly and flatly distributed on all meridian planes 31 of the spherical compound eye structure, the ommatidium optical axes 24 of all ommatidium optical path structures point to the sphere center 32 of the spherical compound eye structure, optical axis included angles 33 between all adjacent ommatidium optical axes 24 are equal, and surface included angles 30 between all adjacent meridian planes 31 are equal.
As shown in fig. 5, the ommatidium light path structure includes an ommatidium optical fiber 3, an ommatidium front lens 25 and an ommatidium rear lens 26, the upper hemispherical compound eye cover 1 and the lower hemispherical compound eye cover 34 are both provided with mounting apertures 23, the ommatidium optical fiber 3 is inserted into the mounting apertures 23 along an ommatidium optical axis 24, the ommatidium front lens 25 is arranged on an end surface of an inner end of the mounting apertures 23, and the ommatidium rear lens 26 is arranged in the middle of the mounting apertures 23.
The output optical fiber 5 and the input optical fiber 19 are connected with the small-eye optical fiber 3 through a Y-shaped joint 4.
N input optical fibers 19 are arranged in the order of the eyelet number and are mounted to the micro lens array 8, and are matched with N small lenses 20; 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 pumping laser for exciting LIF fluorescent signals; the collimating lens 9 expands and collimates the laser 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 fibre 19; the N output optical fibers 5 are arranged according to the sequence of the eyelet numbers and are installed on the photoelectric tube array 6 and are matched with the photodiodes 22 with the same number N; each photodiode 22 covers the band-pass filter 21, the wavelength of the pass band corresponds to the fluorescence band generated by exciting the fluorescence labeling antibody by the pump laser, and the photodiodes 22 convert the fluorescence signals into electric signals; the array driving circuit 7 is used for driving the photoelectric tube array 6 to work, and transmitting the electric signal output by the photoelectric tube array 6 to the main controller 15 for storage and analysis after amplification and analog-to-digital conversion.
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 carry out laser excitation and immunofluorescence receiving, the dynamic detection of the spatial distribution and the change of specific antigen molecules in cells can be realized, and the research requirement on specific physiological movement of the cells is met. Meanwhile, the ommatidium light path structure is arranged on the spherical compound eye structure, so that dynamic detection of cells to be detected in each angle of a three-dimensional space can be realized.
The main controller 15 is used for sending control commands to the laser 10, the leveling controller 35, the flow sample injector 12, the upper head 18, the laser 10, the array driving circuit 7 and the collecting plate 2, controlling the work of the control commands and receiving data of the array driving circuit 7 for storage and analysis.
As shown in fig. 1, 7 and 8, the upper hemispherical compound eye mask body 1 and the lower hemispherical compound eye mask body 34 are provided with two poles 29, and the inner surfaces are formed by metal plating to form inner surface electrodes 36. When the upper hemispherical compound eye mask body 1 and the lower hemispherical compound eye mask body 34 are spliced in a closed manner, 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 sample injector 12 is provided with an upper head 18 for negatively charging the cells to be measured. The flow injector 12 comprises a capillary 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.
Starting the flowing liquid by the flow type sample injector during sample injection, and flowing around the inner periphery of the flow pipe to form sheath flow; the sample cell suspension is ejected from the capillary 37, flows together with the sheath flow 14, forms a single cell flow 16 in the middle of the flow tube, and the outflowing test cell 17 is charged at the outlet of the flow tube 38 by the upper electrode and is 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 cell to be detected with uniform negative charge moves to the condition that the mass center of the cell is approximately superposed with the center of the sphere, and the cell is approximately in a balanced state under the action of electrostatic force, and then the detection can be started; after the detection is finished, the lower hemisphere compound eye and the upper hemisphere compound eye are separated, and the cells to be detected approach the collecting plate under the action of the collecting plate with positive electricity until the collecting plate is attached.
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 carrying out fluorescence labeling on the selected antibody by using a fluorescent agent; directly adding the marked fluorescent antibody on a certain amount of cell samples, dyeing at a certain temperature and time, and washing off the excessive fluorescent antibody which does not participate in the reaction by using water to prepare the sample cell suspension.
And 2, sample introduction, namely adding a sample cell suspension into the spherical compound eye structure in a single cell flow (16) mode, enabling the cell (17) to be detected to carry uniform negative charges through an upper nod, arranging a symmetrical positive electric field in the spherical compound eye structure, and starting detection after the cell (17) to be detected moves until the mass center of the cell is approximately coincident with the sphere center.
Specifically, the step 2 further includes:
starting the flowing liquid by the flow injector (12) to form a sheath flow (14); 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 (38), and the outflowing cell to be tested (17) is electrified by the upper electric head (18) at the outlet of the flow tube (38) and is uniformly negatively charged.
Spherical compound eye structure includes the compound eye cover of upper hemisphere body (34) and the compound eye cover of lower hemisphere body (1), be equipped with the positive electric field of symmetry in the spherical compound eye structure, get into spherical compound eye structure inside back, main control unit (15) send out the instruction and open flat control ware (35), and flat control ware (35) control lower hemisphere compound eye upward movement until the compound eye cover of upper hemisphere body (34) and the compound eye cover of lower hemisphere body (1) amalgamation constitute spherical compound eye structure, and cell (17) that awaits measuring moves to its barycenter and approximately coincides with the centre of sphere, begins to detect this moment.
Specifically, the step 3 further includes:
the main controller (15) starts a laser (10) and an array driving circuit (7), laser emitted by the laser (10) is transmitted along an input optical axis (11), is collimated and expanded by a collimating lens (9), and is focused by each small lens (20) to enter N input optical fibers (19), then enters the small-eye optical fibers (3) in the small-eye optical path structure through a Y-shaped joint (4), and is converged to cells to be detected (17) at the center of the sphere through a small-eye lens group;
the number of the antigen molecules (28) at the intersection point of each ommatidium optical axis (24) and the cell (17) to be detected is different, the fluorescence-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); the distribution of the antigen molecules (28) of the cells to be detected (17) can be inverted by measuring LIF signals generated at the intersection of the optical axis (24) of the ommatidium and the cells to be detected (17);
under the excitation of laser, LIF signals generated at antigen molecules (28) are collected and focused to the end face by each small eye lens group signal, then enter a small eye optical fiber (3), then enter an output optical fiber (5) through a Y-shaped joint (4), the output optical fiber (5) transmits the signals to a photoelectric tube array (6), and a photoelectric diode (22) converts fluorescent signals into electric signals; the array driving circuit (7) amplifies and converts the electric signals output by the photoelectric tube array (6) into analog signals, and then sends the electric signals to the main controller (15) for storage and analysis.
corresponding to LIF data at a certain moment, obtaining the relative number of the antigen molecules (28) at the point by analyzing the intensity of fluorescence at the intersection of each ommatidium optical axis (24) and the cell (17) to be detected, and obtaining the volume distribution of the antigen molecules (28) of the cell (17) to be detected at the moment after calculating all ommatidium on the spherical compound eye structure; the same analysis method is adopted for M moments, and the dynamic change of the distribution of the 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 a three-dimensional space dynamic diagram by system software.
The technical scheme of the invention is based on a bionic compound eye cell detection instrument. The spherical compound eye structure formed by splicing the upper hemispherical compound eye cover body and the lower hemispherical compound eye cover body and the inner surface electrode form an in-sphere uniform electric field, and when the electric field force is balanced, cells which are uniformly charged can be fixed at the center of the sphere, so that the body distribution of the molecules of interest in the cells can be conveniently detected. Each small eye is provided with an LIF light path formed by a small eye light path structure, an excitation light path structure and a fluorescence light path structure to carry out laser excitation and immunofluorescence receiving, so that the dynamic detection of the spatial distribution and the change of specific antigen molecules in cells can be realized, and the research requirement on specific physiological movement of the cells is met. Meanwhile, the ommatidium light path structure is arranged on the spherical compound eye structure, so that dynamic detection of cells to be detected in each angle of a three-dimensional space can be realized. By detecting at fixed time intervals, the dynamic change of the distribution of the antigen molecules of the cells to be detected along with the time can be obtained; the dynamic change can be visually displayed by a three-dimensional space dynamic diagram by system software.
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, selecting a corresponding antibody according to an antigen molecule (28) to be measured, and carrying out fluorescence labeling on the antibody by using a fluorescent agent to prepare a sample cell suspension;
step 2, sample introduction, namely adding a sample cell suspension into a spherical compound eye structure in a single cell flow (16) mode, enabling a cell (17) to be detected to carry uniform negative charges through an upper nod, arranging a symmetrical positive electric field in the spherical compound eye structure, and starting detection after the cell (17) to be detected moves until the mass center of the cell is approximately coincident with the sphere center;
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 excitation light path structure outputs laser to irradiate the cell (17) to be detected through the ommatidium light path structure on the spherical compound eye structure, and LIF signals generated at the intersection of the ommatidium optical axis (24) of the ommatidium light path structure and the cell (17) to be detected are determined through the fluorescence light path structure to invert the volume distribution of the antigen molecules (28) of the cell (17) to be detected;
step 4, LIF signal processing, repeating the step 3 every other fixed time t to obtain a series of LIF signals changing along with time, and finishing detection after the preset detection time number M is finished;
corresponding to LIF data at a certain moment, obtaining the relative number of the antigen molecules (28) at the point by analyzing the intensity of fluorescence at the intersection of each ommatidium optical axis (24) and the cell (17) to be detected, and obtaining the volume distribution of the antigen molecules (28) of the cell (17) to be detected at the moment after calculating all ommatidium on the spherical compound eye structure; the same analysis method is adopted for M moments, and the dynamic change of the distribution of the 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 a three-dimensional space dynamic diagram by system software.
2. The method for detecting somatic cell laser-induced fluorescence based on hemispherical space compound eye-like 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 carrying out fluorescence labeling on the selected antibody by using a fluorescent agent; directly adding the marked fluorescent antibody on a certain amount of cell samples, dyeing at a certain temperature and time, and washing off the excessive fluorescent antibody which does not participate in the reaction by using water to prepare the sample cell suspension.
3. The method for detecting somatic cell laser-induced fluorescence based on hemispherical space compound eye-like structure according to claim 1, wherein the step 2 further comprises:
starting the flowing liquid by the flow injector (12) to form a sheath flow (14); the sample cell suspension is sprayed out of the capillary (37) and flows together with the sheath flow (14) to form a single cell flow (16) in the middle of the flow pipe (38), and the flowing out cell to be detected (17) is electrified by an upper electrode (18) at the outlet of the flow pipe (38) and is uniformly negatively charged;
spherical compound eye structure includes the compound eye cover of upper hemisphere body (34) and the compound eye cover of lower hemisphere body (1), be equipped with the positive electric field of symmetry in the spherical compound eye structure, get into spherical compound eye structure inside back, main control unit (15) send out the instruction and open flat control ware (35), and flat control ware (35) control lower hemisphere compound eye upward movement until the compound eye cover of upper hemisphere body (34) and the compound eye cover of lower hemisphere body (1) amalgamation constitute spherical compound eye structure, and cell (17) that awaits measuring moves to its barycenter and approximately coincides with the centre of sphere, begins to detect this moment.
4. The method for detecting somatic cell laser-induced fluorescence based on hemispherical space compound eye-like structure according to claim 1, wherein the step 3 further comprises:
the main controller (15) starts a laser (10) and an array driving circuit (7), laser emitted by the laser (10) is transmitted along an input optical axis (11), is collimated and expanded by a collimating lens (9), and is focused by each small lens (20) to enter N input optical fibers (19), then enters the small-eye optical fibers (3) in the small-eye optical path structure through a Y-shaped joint (4), and is converged to cells to be detected (17) at the center of the sphere through a small-eye lens group;
the number of the antigen molecules (28) at the intersection point of each ommatidium optical axis (24) and the cell (17) to be detected is different, the fluorescence-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); the distribution of the antigen molecules (28) of the cells to be detected (17) can be inverted by measuring LIF signals generated at the intersection of the optical axis (24) of the ommatidium and the cells to be detected (17);
under the excitation of laser, LIF signals generated at antigen molecules (28) are collected and focused to the end face by each small eye lens group signal, then enter a small eye optical fiber (3), then enter an output optical fiber (5) through a Y-shaped joint (4), the output optical fiber (5) transmits the signals to a photoelectric tube array (6), and a photoelectric diode (22) converts fluorescent signals into electric signals; the array driving circuit (7) amplifies and converts the electric signals output by the photoelectric tube array (6) into analog signals, and then sends the electric signals to the main controller (15) for storage and analysis.
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Citations (21)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US4280623A (en) * | 1980-02-04 | 1981-07-28 | Legorreta Gildardo S | Method and apparatus for controlling, orienting and analyzing biological cells in liquid suspension |
| US5449625A (en) * | 1991-12-20 | 1995-09-12 | Ibiden Co., Ltd. | Optical fiber based fluorescent immunoassay apparatus |
| JPH08124828A (en) * | 1994-10-24 | 1996-05-17 | Nec Corp | Exposure light projection device |
| US5926271A (en) * | 1995-12-20 | 1999-07-20 | Zeta Technology | Laser-induced fluorescence detector having a capillary detection cell and method for identifying trace compounds implemented by the same device |
| JP2000241428A (en) * | 1999-02-19 | 2000-09-08 | Japan Advanced Inst Of Science & Technology Hokuriku | Method and device for measuring antigen |
| JP2001117187A (en) * | 1999-10-15 | 2001-04-27 | Hiroshi Takoi | Full-space photographic compound eye camera |
| US20030054569A1 (en) * | 2001-09-06 | 2003-03-20 | Cheng Anthony K. | Particle based homogeneous assays using capillary electrophoresis with laser-induced fluorescence detection |
| CN1712939A (en) * | 2004-06-22 | 2005-12-28 | 中国科学院大连化学物理研究所 | Cell imaging method based on laser induced fluorescence |
| US20060119845A1 (en) * | 2004-12-07 | 2006-06-08 | Leonard Stephen W | Optical system |
| JP2007117234A (en) * | 2005-10-26 | 2007-05-17 | Graduate School For The Creation Of New Photonics Industries | Anticaries and diagnostic apparatus |
| US7286295B1 (en) * | 2005-11-30 | 2007-10-23 | Sandia Corporation | Microoptical compound lens |
| CN101105455A (en) * | 2007-07-04 | 2008-01-16 | 四川大学 | Laser induction fluorescence detector |
| CN102033124A (en) * | 2009-12-25 | 2011-04-27 | 北京博晖创新光电技术股份有限公司 | Immunofluorescence detection device and detection method |
| US20120007815A1 (en) * | 2010-07-09 | 2012-01-12 | Samsung Electronics Co., Ltd. | Multipurpose sensing apparatus and electronic equipment having the same |
| US20140014855A1 (en) * | 2012-07-12 | 2014-01-16 | KAIST (Korea Advanced Institute of Science and Technology) | Condensing-type portable fluorescence detection system |
| WO2016148210A1 (en) * | 2015-03-18 | 2016-09-22 | コニカミノルタ株式会社 | Light source device and projection device |
| CN106526606A (en) * | 2016-10-13 | 2017-03-22 | 中国科学院上海技术物理研究所 | Double-channel smart quantum dot laser space trajectory detection system based on bionic compound eyes |
| CN206348269U (en) * | 2016-10-27 | 2017-07-21 | 西安精英光电技术有限公司 | A kind of bioluminescence collection structure based on ellipsoidal mirror |
| KR101820632B1 (en) * | 2017-07-12 | 2018-01-19 | 레이콜 | Lens provided with compound eye structure and camera system including it |
| AU2020101129A4 (en) * | 2020-06-25 | 2020-07-30 | Guilin Univ. Electr. Techol. | A microsphere self-assembled laser system based on a coaxial three-waveguide optical fiber |
| CN111487233A (en) * | 2020-05-27 | 2020-08-04 | 安徽皖仪科技股份有限公司 | PCR light path system based on fly-eye lens |
-
2021
- 2021-03-15 CN CN202110277220.9A patent/CN113063759B/en active Active
Patent Citations (21)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US4280623A (en) * | 1980-02-04 | 1981-07-28 | Legorreta Gildardo S | Method and apparatus for controlling, orienting and analyzing biological cells in liquid suspension |
| US5449625A (en) * | 1991-12-20 | 1995-09-12 | Ibiden Co., Ltd. | Optical fiber based fluorescent immunoassay apparatus |
| JPH08124828A (en) * | 1994-10-24 | 1996-05-17 | Nec Corp | Exposure light projection device |
| US5926271A (en) * | 1995-12-20 | 1999-07-20 | Zeta Technology | Laser-induced fluorescence detector having a capillary detection cell and method for identifying trace compounds implemented by the same device |
| JP2000241428A (en) * | 1999-02-19 | 2000-09-08 | Japan Advanced Inst Of Science & Technology Hokuriku | Method and device for measuring antigen |
| JP2001117187A (en) * | 1999-10-15 | 2001-04-27 | Hiroshi Takoi | Full-space photographic compound eye camera |
| US20030054569A1 (en) * | 2001-09-06 | 2003-03-20 | Cheng Anthony K. | Particle based homogeneous assays using capillary electrophoresis with laser-induced fluorescence detection |
| CN1712939A (en) * | 2004-06-22 | 2005-12-28 | 中国科学院大连化学物理研究所 | Cell imaging method based on laser induced fluorescence |
| US20060119845A1 (en) * | 2004-12-07 | 2006-06-08 | Leonard Stephen W | Optical system |
| JP2007117234A (en) * | 2005-10-26 | 2007-05-17 | Graduate School For The Creation Of New Photonics Industries | Anticaries and diagnostic apparatus |
| US7286295B1 (en) * | 2005-11-30 | 2007-10-23 | Sandia Corporation | Microoptical compound lens |
| CN101105455A (en) * | 2007-07-04 | 2008-01-16 | 四川大学 | Laser induction fluorescence detector |
| CN102033124A (en) * | 2009-12-25 | 2011-04-27 | 北京博晖创新光电技术股份有限公司 | Immunofluorescence detection device and detection method |
| US20120007815A1 (en) * | 2010-07-09 | 2012-01-12 | Samsung Electronics Co., Ltd. | Multipurpose sensing apparatus and electronic equipment having the same |
| US20140014855A1 (en) * | 2012-07-12 | 2014-01-16 | KAIST (Korea Advanced Institute of Science and Technology) | Condensing-type portable fluorescence detection system |
| WO2016148210A1 (en) * | 2015-03-18 | 2016-09-22 | コニカミノルタ株式会社 | Light source device and projection device |
| CN106526606A (en) * | 2016-10-13 | 2017-03-22 | 中国科学院上海技术物理研究所 | Double-channel smart quantum dot laser space trajectory detection system based on bionic compound eyes |
| CN206348269U (en) * | 2016-10-27 | 2017-07-21 | 西安精英光电技术有限公司 | A kind of bioluminescence collection structure based on ellipsoidal mirror |
| KR101820632B1 (en) * | 2017-07-12 | 2018-01-19 | 레이콜 | Lens provided with compound eye structure and camera system including it |
| CN111487233A (en) * | 2020-05-27 | 2020-08-04 | 安徽皖仪科技股份有限公司 | PCR light path system based on fly-eye lens |
| AU2020101129A4 (en) * | 2020-06-25 | 2020-07-30 | Guilin Univ. Electr. Techol. | A microsphere self-assembled laser system based on a coaxial three-waveguide optical fiber |
Non-Patent Citations (2)
| Title |
|---|
| 郝永平等: "仿生复眼结构设计及其成像系统研究新进展" * |
| 雷卫宁等: "基于仿生复眼的大视场探测系统结构研究", 《光学与光电技术》 * |
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