High-throughput fluorescence mapping system based on imaging fiber array

Abstract

The application discloses a high-throughput fluorescence mapping system based on an imaging fiber array and relates to the technical field of electrophysiological experimental equipment. The application discloses a high-throughput fluorescence mapping system based on an imaging fiber array and relates to the technical field of electrophysiological experimental equipment. The application discloses a high-throughput fluorescence mapping system based on an imaging fiber array and relates to the technical field of electrophysiological experimental equipment. The application discloses a high-throughput fluorescence mapping system based on an imaging fiber array and relates to the technical field of electrophysiological experimental equipment. The application discloses a high-throughput fluorescence mapping system based on an imaging fiber array and relates to the technical field of electrophysiological experimental equipment. The application discloses a high-throughput fluorescence mapping system based on an imaging fiber array and relates to the technical field of electrophysiological experimental equipment. The application discloses a high-throughput fluorescence mapping system based on an imaging fiber array and relates to the technical field of electrophysiological experimental equipment. The application discloses a high-throughput fluorescence mapping system based on an imaging fiber array and relates to the technical field of electrophysiological experimental equipment. The application discloses a high-throughput fluorescence mapping system based on an imaging fiber array and relates to the technical field of electrophysiological experimental equipment. The application discloses a high-throughput fluorescence mapping system based on an imaging fiber array and relates to the technical field of electrophysiological experimental equipment. The application discloses a high-throughput fluorescence mapping system based on an imaging fiber array and relates to the technical field of electrophysiological experimental equipment. The application discloses a high-throughput fluorescence mapping system based on an imaging fiber array and relates to the technical field of electrophysiological experimental equipment. The application discloses a high-throughput fluorescence mapping system based on an imaging fiber array and relates to the technical field of electrophysiological experimental equipment. The application discloses a high-throughput fluorescence mapping system based on an imaging fiber array and relates to the technical field of electrophysiological experimental equipment. The application discloses a high-throughput fluorescence mapping system based on an imaging fiber array and relates to the technical field of electrophysiological experimental equipment. The application discloses a high-throughput fluorescence mapping system based on an imaging fiber array and relates to the technical field of electrophysiological experimental equipment. The application discloses a high-throughput fluorescence mapping

Description

Technical Field

[0001] The present invention relates to the technical field of fluorescence mapping electrophysiological experimental equipment, and in particular to a high-throughput fluorescence mapping system based on an imaging optical fiber array. Background Art

[0002] In fluorescence mapping electrophysiological experiments, it is sometimes necessary to perform a large number of repeated experiments on cells using cell culture plates to obtain batch data for analysis. Given the huge volume of such experimental data, manual processing is time-consuming and labor-intensive, and may even introduce more additional interference, resulting in inconsistency in the experimental data, making it difficult to achieve the experimental purpose.

[0003] Currently, the equipment on the market mainly uses LED light sources to directly excite the cells in each culture well at the bottom of the cell culture well plate. Since the area of ​​the bottom of the cell culture well plate is relatively large, there will inevitably be differences in the excitation light sources obtained between the various culture wells.

[0004] In addition, the imaging process also involves directly capturing the entire cell at the bottom of the cell culture plate through a camera. This is also limited by the relatively large area of ​​the bottom of the cell culture plate, resulting in a relatively long imaging distance after focusing, and the weakness of the fluorescence signal itself, which further attenuates the light intensity reaching the camera. In addition, since the fluorescence signal is easily affected by stray light during its long-distance propagation, it is difficult to ensure the consistency of cell imaging between each well of the cell culture plate.

[0005] Although there are products in the existing technology that combine optical fibers and microscopes for cell imaging, they are limited by the microscope's own functional application for magnification, and cannot complete large-area data acquisition, and thus cannot achieve high-throughput imaging of multi-channel fluorescence signals.

[0006] In addition, it is difficult to strictly control the temperature, humidity and gas concentration in the space during the experiment with existing technologies, resulting in experimental results that cannot meet the requirements. Summary of the Invention

[0007] The technical problem to be solved by the present invention is to overcome the existing defects and provide a high-throughput fluorescence mapping system based on an imaging fiber array to solve the problems in the prior art, such as uneven excitation light received by each culture well due to the large bottom area of ​​the cell culture well plate, increased attenuation of the fluorescence signal during propagation due to the relatively large bottom area of ​​the cell culture well plate and the relatively long imaging shooting distance, as well as the susceptibility to stray light during propagation, resulting in inconsistent cell imaging effects between each well of the cell culture well plate.

[0008] To achieve the above objectives, the present invention provides the following technical solution: a high-throughput fluorescence mapping system based on an imaging fiber array, comprising a housing, a clamping module, an imaging module, and a cell culture well plate, wherein an operating table is provided in the middle portion of the inner side of the housing, and the operating table divides the interior space of the housing into an upper and lower portion, wherein the upper portion of the operating table is a cell manipulation space, and the lower portion of the operating table is a cell imaging space, and a plate slot is provided on the top of the operating table;

[0009] The clamping module is arranged in the cell operation space, and the clamping module includes a longitudinal electric slide rail and a transverse electric slide rail. The longitudinal electric slide rail is provided with two respectively arranged on both sides of the inner top surface of the box body, and the top ends of the transverse electric slide rail are respectively fixedly connected to the sliding parts of the longitudinal electric slide rails on both sides. The bottom of the sliding part of the transverse electric slide rail is provided with an electric telescopic rod, and the bottom end of the electric telescopic rod is provided with a frame seat. The bottom of the frame seat is slidably connected to the slide rail, and the interior of the slide rail is rotatably connected to a bidirectional screw, and both sides of the bidirectional screw are threadedly sleeved with a clamping plate;

[0010] The imaging module is arranged in the cell imaging space, and comprises a sealing cover, an imaging optical fiber array, a focusing module, a light splitting module, a light source module, an imaging camera and an imaging optical fiber array lens.

[0011] Furthermore, the imaging fiber array is an array composed of a plurality of imaging optical fibers, one end of the imaging fiber array is gathered and the other end is dispersed, the imaging fiber array lens is arranged in a rectangular array at the bottom of the middle plate slot, the sealing cover is arranged at the bottom of the operating table at a position corresponding to the middle plate slot, the top of the imaging fiber array passes through the bottom of the sealing cover, the spectrometer module is arranged at the bottom end of the imaging fiber array, the light source module is arranged on one side of the spectrometer module, and the imaging camera is arranged on the outer surface of the spectrometer module;

[0012] The outside of the imaging optical fiber array is fixedly sleeved with a limiting cylinder, the outside of the limiting cylinder is provided with a support, and the support is provided on the inner bottom surface of the box body;

[0013] The imaging fiber array lenses correspond one-to-one to the bottom of the wells in the cell culture well plate. The optical fibers at one end of the imaging fiber array located inside the sealing cover are divergently connected to the bottom of each imaging fiber array lens. The focusing module is arranged between the imaging fiber array and the spectroscopic module.

[0014] Furthermore, a dichroic mirror and a filter are provided inside the light splitting module to separate the light of different wavelengths;

[0015] The imaging camera is a high-speed camera with a shooting rate greater than 400 FPS to capture changes in high-speed fluorescence signals. Several imaging cameras are installed outside the spectroscopic module to simultaneously record and collect various fluorescence images.

[0016] Furthermore, threads with opposite spiral directions are symmetrically provided on both sides of the bidirectional screw, the top of the clamping plate is slidably matched with the slide rail, magnetic blocks are provided at both ends of the slide rail, a clamping motor is provided at one end of the slide rail, the output shaft of the clamping motor is fixedly connected to the end of the bidirectional screw, and clamping seats are symmetrically provided at the bottom ends of the clamping plates on both sides, and arc-shaped grooves are provided on opposite sides of the clamping seats on both sides;

[0017] A rotary motor is embedded in the bottom of the sliding member of the transverse electric slide rail, and the output shaft of the rotary motor is fixedly connected to the fixed end of the electric telescopic rod.

[0018] Furthermore, a tooth plate is provided on the top of the slide rail, and a transmission gear is rotatably connected to the interior of the frame seat through a rotating shaft, and the transmission gear is engaged with the tooth plate. A drive motor is provided on the side of the frame seat, and the output shaft of the drive motor is fixedly connected to the end of the rotating shaft.

[0019] Furthermore, a side groove is provided on the side of the box at a position corresponding to the cell operation space, a door panel is movably provided on one side of the side groove, a temperature and humidity sensor and a gas sensor are provided on the door panel, one side of the box is transparent, and a sealing door is movably provided on the transparent side of the box;

[0020] There are five plate slots, and the bottom of the plate slot located in the middle is a hollow structure. A cell culture well plate is placed in the plate slot.

[0021] Furthermore, a temperature control module is provided on the top of the box body, and the temperature control module includes a heat pipe, a heat insulation frame and a liquid tank. The heat pipe is provided on the top side of the box body, and the two ends of the heat pipe are respectively provided with a liquid inlet pipe and a liquid return pipe that pass through the top surface of the box body. The bottom surface of the heat pipe is provided with a heat spreader, and curved plates are symmetrically provided on both sides of the heat spreader, and magnetic strips are embedded on the curved side surfaces of the curved plates.

[0022] The insulation frame is arranged on the top surface of the box body, and the top side of the insulation frame is rotatably connected to a thermoelectric cooling plate. The bottom of the insulation frame is horizontally penetrated by heat exchange tubes in a linear array, and the two ends of the heat exchange tubes are respectively provided with a collecting pipe and a diverter pipe;

[0023] The liquid tank is arranged on the top surface of the box body, and a circulation pump is provided on the liquid tank.

[0024] Furthermore, the heat pipe is arranged in an S shape, the arc-shaped plate is made of elastic material, a flip motor is provided on the side of the heat preservation frame, and the output shaft of the flip motor is fixedly connected to the middle of the end surface of the thermoelectric cooling plate;

[0025] The collecting pipe is fixedly connected to the top end of the liquid inlet pipe;

[0026] The input port of the circulation pump is fixedly connected to the liquid tank, and the output port of the circulation pump is fixedly connected to the diversion pipe through a hose.

[0027] Furthermore, a gas regulating module is provided on the top of the box body, and the gas regulating module includes a gas supply tank group, a water tank and an electric push rod. The water tank is provided on the top surface of the box body, and a water pump is provided on one side of the water tank;

[0028] The electric push rods are symmetrically arranged in two groups, each group contains two, and the two groups of electric push rods are symmetrically arranged on both sides of the top surface of the box body. An introduction pipe is provided between the bottom telescopic ends of the two electric push rods in the same group, and nozzles are evenly arranged on the introduction pipe.

[0029] Furthermore, the gas supply tank group includes several types of gas compression tanks, and the tops of each gas compression tank in the gas supply tank group are connected to a gas collecting pipe through a solenoid valve. The gas collecting pipe is provided with an air guide pipe, the input port of the water pump is fixedly connected to the water tank, and the output port of the water pump is provided with a flow guide pipe. One end of the inlet pipes on both sides are fixedly connected to the air guide pipe and the flow guide pipe respectively, and the middle part of the inlet pipes on both sides is fixedly sleeved with an adjusting gear.

[0030] Compared with the prior art, the present invention has the following advantages:

[0031] 1. The present invention cultures cells in a cell culture well plate. The fluorescent signal generated by the excitation is transmitted through a specific imaging fiber array at the bottom of the cell culture well plate, and finally reaches an imaging camera for imaging after passing through a spectrometer, thereby realizing batch collection of experimental data and avoiding signal attenuation and interference from stray light during the transmission process. The excitation light of the light source module passes through the spectrometer and the imaging fiber array and directly reaches the bottom of the cell culture well plate to excite the cells containing the fluorescent dye, thereby avoiding the problem of uneven excitation light intensity received by each culture well.

[0032] 2. When humidifying or filling the cell operation space with gas, the present invention uses a rotating motor to make the slide rail perpendicular to the horizontal electric slide rail, and then uses a driving motor to drive the slide rail to move horizontally for a certain distance, and then the electric push rod drives the introduction tube to move downward, so that the adjustment gear engages with the gear plate, and then the driving motor is rotated back and forth to cause the introduction tube to rotate back and forth, so that the humidified airflow or the added gas is evenly distributed in the cell operation space.

[0033] 3. In the present invention, after the adjustment gear is engaged with the gear plate, the nozzle is rotated to correspond to the bottom surface of the heat spreader, and then the drive motor is used to swing the inlet tube back and forth. The humidified airflow or gas ejected from the nozzle reaches the same or similar temperature as the cell operation space after contacting the heat spreader. When the inlet tube swings back and forth, the humidified airflow or gas ejected is temperature-compensated and then flows along the arc plate to the plate position slot, thereby preventing the humidified airflow or gas from causing temperature fluctuations in the cell operation space. BRIEF DESCRIPTION OF THE DRAWINGS

[0034] Figure 1 It is a schematic diagram of the structure of the present invention;

[0035] Figure 2 This is a structural schematic diagram of the present invention from another angle;

[0036] Figure 3 Schematic diagram of the internal structure of the present invention;

[0037] Figure 4 It is a schematic diagram of the front view structure of the present invention;

[0038] Figure 5 This is a schematic diagram of the structure of the present invention when viewed from above;

[0039] Figure 6 For the present invention Figure 5 A in the middle is an enlarged structural diagram;

[0040] Figure 7 It is a schematic cross-sectional view of the present invention;

[0041] Figure 8 For the present invention Figure 7 Enlarged structural diagram at point B in the middle.

[0042] In the figure: 1. Box body; 101. Operation table; 102. Side groove; 103. Door panel; 104. Sealed door; 105. Plate slot; 2. Clamping module; 201. Longitudinal electric slide rail; 202. Horizontal electric slide rail; 203. Electric telescopic rod; 204. Frame seat; 205. Slide rail; 206. Bidirectional screw; 207. Clamping plate; 208. Clamping seat; 209. Arc groove; 210. Tooth plate; 211. Rotating shaft; 212. Transmission gear; 213. Driving motor; 214. Rotating motor; 215. Clamping motor; 3. Imaging module; 301. Sealing cover; 302. Imaging fiber array; 303. Limiting cylinder; 304. Support; 305. Focusing module; 306. Spectrometer module; 307. Light source module; 308. Imaging camera; 309. Imaging fiber array lens; 4. Temperature control module; 401. Heat pipe; 402. Liquid inlet pipe; 403. Liquid return pipe; 404. Insulation frame; 405. Thermoelectric cooling plate; 406. Flip motor; 407. Heat exchange tube; 408. Liquid tank; 409. Circulation pump; 410. Heat spreader; 411. Curved plate; 412. Magnetic strip; 5. Gas regulating module; 501. Gas supply tank group; 502. Gas collecting pipe; 503. Gas guide pipe; 504. Water tank; 505. Water pump; 506. Flow guide pipe; 507. Electric push rod; 508. Inlet pipe; 509. Nozzle; 510. Adjustment gear. DETAILED DESCRIPTION

[0043] The following will clearly and completely describe the technical solutions in the embodiments of the present invention in conjunction with the accompanying drawings. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without making creative efforts are within the scope of protection of the present invention.

[0044] Example 1

[0045] See also Figure 1-8 The present embodiment provides a high-throughput fluorescence mapping system based on an imaging fiber array, including a box body 1, a clamping module 2 and an imaging module 3. An operating table 101 is provided in the middle of the inner side of the box body 1. The operating table 101 is used to carry a cell culture well plate. A plate slot 105 is provided on the top of the operating table 101. There are five plate slots 105. The bottom of the plate slot 105 located in the middle is a hollow structure. Cell culture well plates are placed in the plate slots 105. The cell culture well plates are made of transparent material, and a pipette cleaning slot is placed in the last plate slot 105 for cleaning the tip of the pipette.

[0046] like Figure 3 As shown, the five plate slots 105 are counted as the first plate position, the second plate position, the third plate position, the fourth plate position and the fifth plate position from left to right. The first plate position is used for parking and adding liquid to the cell culture well plate entering the cell operation space. At the same time, a heating module is integrated to heat the bottom of the cell culture well plate. This plate position can also be used as a parking position for replacing the pipette tip. The unloaded pipette tip is moved out of the cell operation space by the clamping module 2 at this plate position, and the pipette tip moved into the cell operation space by the clamping module 2 is loaded at this plate position; the second plate position is used for parking and adding liquid to the cell culture well plate to be imaged. This plate position is also integrated with a heating module. The bottom of the cell culture well plate is heated; the third plate position is used to park the imaging cell culture well plate, and the bottom of the plate position is hollowed out to facilitate signal acquisition and optical imaging at the diverging end of the imaging fiber array 302. This plate position is integrated with an electrical stimulation module to facilitate electrical stimulation of cells according to experimental requirements; the fourth plate position is used to park and add liquid to the cell culture well plate that has completed imaging or needs to be imaged again. It can also be used as a parking position for cell containers for certain specific experiments. This plate position is also integrated with a heating function to heat the bottom of the cell culture well plate or cell container; the fifth plate position is used to park the pipette cleaning tank to clean the pipette tip.

[0047] The operating table 101 divides the internal space of the box 1 into two parts, the upper part of the operating table 101 is the cell operation space, which mainly includes the operating table 101, the clamping module 2, the pipette (not shown in the figure), the cell culture well plate, the cell container (not shown in the figure), the pipette cleaning tank, etc.

[0048] The lower part of the operating table 101 is a cell imaging space, which includes an imaging module 3. The imaging module 3 includes a sealing cover 301, an imaging fiber array 302, a focusing module 305, a spectroscopic module 306, a light source module 307, an imaging camera 308 and an imaging fiber array lens 309. The sealing cover 301 is arranged at the bottom of the operating table 101 and at a position corresponding to the middle plate slot 105. The sealing cover 301 is used to seal the divergent end of the imaging fiber array 302 to prevent interference from stray light.

[0049] The top of the imaging fiber array 302 passes through the bottom of the sealing cover 301. The imaging fiber array 302 is an array composed of a plurality of imaging optical fibers, with one end of the imaging fiber array 302 being concentrated and the other end being dispersed.

[0050] Specifically, the imaging fiber has an imaging function compared to the ordinary lighting fiber. The imaging fiber forms an image relative to the lens, has the advantage of less light loss, and extends the optical path distance.

[0051] Compared with lens imaging, the imaging fiber array has more flexible shooting positions and more uniform excitation light. It is especially suitable for multi-well plate imaging and greatly reduces the invalid imaging area.

[0052] The light splitting module 306 is disposed at the bottom end of the imaging fiber array 302 . A dichroic mirror and a filter are disposed inside the light splitting module 306 to separate light of different wavelengths.

[0053] The focusing module 305 is disposed between the imaging fiber array 302 and the light splitting module 306 .

[0054] The light source module 307 is disposed on one side of the light splitting module 306 .

[0055] The imaging camera 308 is arranged on the outer surface of the spectroscopic module 306. The imaging camera 308 is a high-speed camera with a shooting rate greater than 400 FPS, preferably a high-speed sCMOS camera, to capture changes in high-speed fluorescence signals. Several imaging cameras 308 are installed on the outside of the spectroscopic module 306 to simultaneously record and collect various fluorescence images.

[0056] The imaging fiber optic array lens 309 is arranged in a rectangular array at the bottom of the middle plate slot 105. The imaging fiber optic array lens 309 corresponds one-to-one to the bottom of the hole in the cell culture well plate. The optical fiber at one end of the imaging fiber optic array 302 located inside the sealing cover 301 is divergent and connected to the bottom of each imaging fiber optic array lens 309. The outside of the imaging fiber optic array 302 is fixedly sleeved with a limiting cylinder 303, and a support 304 is provided on the outside of the limiting cylinder 303. The support 304 is arranged on the inner bottom surface of the box body 1.

[0057] Specifically, the specifications and models of the imaging fiber optic array 302 are strictly adapted to the cell culture well plate, and multiple configurations such as 6 bundles, 12 bundles, 24 bundles, 48 ​​bundles, 96 bundles, and 384 bundles are provided. Before each experiment, the imaging fiber optic array 302 corresponding to the channel needs to be installed according to the specifications of the cell culture well plate. The signal acquisition end (i.e., the divergent end) of the imaging fiber optic array 302 is located at the bottom of the cell culture well plate. There is an optical fiber under each culture well of the cell culture well plate. The imaging end of the imaging fiber optic array 302 is facing the high-speed imaging lens on the spectrometer module 306. One or more high-speed sCMOS cameras can be connected to the spectrometer module 306. The spectrometer module 306 is a fluorescence beam splitter, and the light source module 307 is preferably an LED excitation light source.

[0058] A side groove 102 is provided on the side of the box body 1 at a position corresponding to the cell operation space. A door panel 103 is movably provided on one side of the side groove 102. A temperature and humidity sensor and a gas sensor are provided on the door panel 103. The temperature and humidity sensor is used to detect the temperature and humidity in the cell operation space, and the gas sensor is used to detect the gas concentration in the cell operation space. The side groove 102 facilitates the entry and exit of experimental equipment or materials. One side of the box body 1 is transparent, and a sealing door 104 is movably provided on the transparent side of the box body 1. The door panel 103 and the sealing door 104 are preferably electric doors for easy automatic opening and closing.

[0059] The clamping module 2 is arranged in the cell operation space. The clamping module 2 includes a longitudinal electric slide 201 and a transverse electric slide 202. The longitudinal electric slide 201 has two respectively arranged on both sides of the inner top surface of the box body 1. The top two ends of the transverse electric slide 202 are respectively fixedly connected to the sliding parts of the longitudinal electric slide 201 on both sides. The longitudinal electric slide 201 and the transverse electric slide 202 are respectively used to realize the longitudinal and transverse movement of the clamping seat 208, so as to facilitate the control of the position of the clamping seat 208 in the cell operation space.

[0060] An electric telescopic rod 203 is provided at the bottom of the sliding part of the horizontal electric slide rail 202. The electric telescopic rod 203 is used to realize the lifting and lowering of the clamping seat 208. A frame seat 204 is provided at the bottom end of the electric telescopic rod 203. The bottom of the frame seat 204 is slidably connected to the slide rail 205. The internal rotation of the slide rail 205 is connected to a bidirectional screw 206. The two sides of the bidirectional screw 206 are symmetrically provided with threads with opposite spiral directions. Both sides of the bidirectional screw 206 are threadedly sleeved with clamping plates 207. The top of the clamping plate 207 slides with the slide rail 205. The bottom ends of the clamping plates 207 on both sides are symmetrically provided with clamping seats 208. A clamping motor 215 is provided at one end of the slide rail 205. The output shaft of the clamping motor 215 is fixedly connected to the end of the bidirectional screw 206.

[0061] Specifically, the clamping motor 215 drives the bidirectional screw 206 to rotate, so that the clamping plates 207 on both sides drive the clamping seat 208 to move in opposite directions, so as to achieve clamping by using the end surface of the clamping seat 208.

[0062] The opposite sides of the clamping seats 208 on both sides are provided with arc-shaped grooves 209. The arrangement of the arc-shaped grooves 209 makes it easier for the clamping seats 208 to clamp the pipette and facilitate pipetting.

[0063] A rotating motor 214 is embedded in the bottom of the sliding part of the horizontal electric slide 202. The output shaft of the rotating motor 214 is fixedly connected to the fixed end of the electric telescopic rod 203. The rotating motor 214 is used to drive the electric telescopic rod 203 to rotate and drive the clamping seat 208 at the bottom to rotate, so as to facilitate the adjustment of the angle of the clamped cell culture well plate, pipette and other experimental equipment.

[0064] A tooth plate 210 is provided on the top of the slide rail 205, and a transmission gear 212 is rotatably connected to the interior of the frame seat 204 through a rotating shaft 211. The transmission gear 212 is engaged with the tooth plate 210. A drive motor 213 is provided on the side of the frame seat 204. The output shaft of the drive motor 213 is fixedly connected to the end of the rotating shaft 211. The drive motor 213 drives the transmission gear 212 to rotate through the rotating shaft 211. The transmission gear 212 realizes the horizontal linear motion of the slide rail 205 by engaging with the tooth plate 210, which is used to clamp experimental equipment and other materials in and out of the cell operation space.

[0065] A temperature control module 4 is provided on the top of the box body 1. The temperature control module 4 includes a heat pipe 401, an insulation frame 404 and a liquid tank 408. The heat pipe 401 is provided on the internal top side of the box body 1. The heat pipe 401 is arranged in an S shape. The two ends of the heat pipe 401 are respectively provided with a liquid inlet pipe 402 and a liquid return pipe 403 that pass through the top surface of the box body 1. A heat spreader 410 is provided on the bottom surface of the heat pipe 401. The heat spreader 410 is made of a heat-conducting material. The heat spreader 410 is used to exchange heat with the medium in the heat pipe 401 to regulate the temperature in the cell operation space.

[0066] The insulation frame 404 is arranged on the top surface of the box body 1. The top side of the insulation frame 404 is rotatably connected to the thermoelectric cooling piece 405. The thermoelectric cooling piece 405 is a semiconductor cooling piece. The side of the insulation frame 404 is provided with a flip motor 406. The output shaft of the flip motor 406 is fixedly connected to the middle of the end face of the thermoelectric cooling piece 405. The flip motor 406 drives the thermoelectric cooling piece 405 to rotate 180 degrees to realize the flip of the thermoelectric cooling piece 405, thereby realizing the hot end and the cold end of the thermoelectric cooling piece 405. Switch, the bottom of the insulation frame 404 is horizontally penetrated by heat exchange tubes 407 in a linear array. The heat exchange tubes 407 are arranged in a linear array to facilitate rapid heat exchange with the flowing medium in the heat exchange tubes 407. The two ends of the heat exchange tubes 407 are respectively provided with a collecting pipe and a diverter pipe. The collecting pipe is fixedly connected to the top of the liquid inlet pipe 402. When the hot end of the thermoelectric cooling plate 405 is flipped to face the inside of the insulation frame 404, the heat exchange tubes 407 passing through the insulation frame 404 and the medium flowing therein are heated.

[0067] The liquid tank 408 is arranged on the top surface of the box body 1. The liquid tank 408 contains a flowing medium. A circulating pump 409 is provided on the liquid tank 408. The input port of the circulating pump 409 is fixedly connected to the liquid tank 408. The output port of the circulating pump 409 is fixedly connected to the diversion pipe through a hose.

[0068] A gas regulating module 5 is provided on the top of the box body 1. The gas regulating module 5 includes a gas supply tank group 501, a water tank 504 and an electric push rod 507. The gas supply tank group 501 includes several gas compression tanks for storing gases required for cell culture, such as carbon dioxide and oxygen. The tops of the various gas compression tanks in the gas supply tank group 501 are connected to a gas collecting pipe 502 through a solenoid valve. The gas collecting pipe 502 is provided with an air guide pipe 503. The water tank 504 is provided on the top surface of the box body 1. The water tank 504 contains sterile liquid for humidification. The water tank 504 is a sterile water tank. A water pump 505 is provided on one side of the water tank 504. The input port of the water pump 505 is fixedly connected to the water tank 504, and the output port of the water pump 505 is provided with a guide pipe 506. Two groups of electric push rods 507 are symmetrically provided, each group contains two, and the two groups of electric push rods 507 are symmetrically arranged on both sides of the top surface of the box body 1. An inlet pipe 508 is provided between the bottom telescopic ends of the two electric push rods 507 in the same group, and nozzles 509 are evenly provided on the inlet pipe 508. One end of the inlet pipes 508 on both sides is fixedly connected to the air guide pipe 503 and the guide pipe 506 respectively. The nozzle 509 connected to the air supply pipeline is an air nozzle, and the nozzle 509 connected to the liquid pipeline is an atomizing nozzle.

[0069] The cam 206 is rotated to move the lever 208 toward the bottom of the drawer 202, and the lever 208 is moved to the bottom of the drawer 202. The lever 206 is rotated to move the lever 208 toward the bottom of the drawer 202. The rail 202 moves the clamped cell culture well plate into the cell operation space, and then uses the downward extension of the electric telescopic rod 203 to place the cell culture well plate in the corresponding plate slot 105 and release the cell culture well plate to reset the clamping seat 208. Next, the clamping seat 208 is moved to the position corresponding to the pipette, so that the arc groove 209 corresponds to the side of the pipette and the clamping operation of the pipette is completed by closing the arc groove 209 in the above manner. The pipette is preferably an electric pipette to facilitate automatic extraction and discharge. The pipette is then driven to collect cells from the cell container and inject them one by one into the culture wells of the cell culture well plate, and the heating module and the electric stimulation module on the plate slot 105 are started according to experimental needs. After the liquid is added to the cell culture well plate, the clamping module 2 is used to place the cell culture well plate after adding liquid in the middle plate slot 105. At this time, each imaging fiber array lens 309 corresponds to the culture well of the cell culture well plate.

[0070] During the experiment, the LED excitation light generated by the light source module 307 enters the imaging fiber array 302 through the high-speed lens on the spectrometer module 306. Utilizing the characteristic that fluorescent substances emit fluorescence when excited by light of a specific wavelength, the excitation light is transmitted to the culture wells of the cell culture well plate through the imaging fiber array 302, thereby exciting the fluorescent substances in the samples in the culture wells to generate fluorescence. The generated fluorescence signal then enters the spectrometer module 306 through the high-speed lens, and imaging is achieved using the imaging camera 308 provided on the spectrometer module 306.

[0071] More specifically, the spectrometer module 306 has a high-quality fluorescence filter set inside. These filters can be flexibly configured according to experimental requirements. The LED excitation light source has a variety of bands to choose from. The single-band or multi-band excitation light required by the fluorescent dye is transmitted into the optical fiber in a unidirectional manner through the spectrometer module 306, and finally reaches the bottom of the cell culture well plate to excite the cells fluorescently. The spectrometer module 306 also divides the fluorescence signal to be collected into different optical paths for transmission, and finally the fluorescence of each band reaches the corresponding imaging camera 308 for imaging.

[0072] More specifically, in order to achieve consistency in the signal of each optical fiber, the present mapping system is provided with a fiber combiner at the signal acquisition end and the imaging end of the imaging fiber array 302, which have various specifications such as 6 holes, 12 holes, 24 holes, 48 ​​holes, 96 holes, and 384 holes. The fiber combiner at the acquisition end of the imaging fiber array 302 aligns and clamps the optical fibers in the imaging fiber array 302 according to the specifications of the cell culture well plate. The spacing between each optical fiber hole of the fiber combiner is completely consistent with the cell culture well plate, so that all optical fiber acquisition planes inserted into the fiber combiner are at the same level, and each optical fiber is facing the bottom of a culture well. The fiber combiner at the imaging end of the imaging fiber array 302 makes the optical fibers more concentrated and controls all the optical fiber imaging end planes on the same horizontal plane, thereby improving the quality of fluorescence imaging and further enhancing the consistency of signals between channels.

[0073] Furthermore, this mapping system can also configure a miniature high-speed lens for each optical fiber of the imaging fiber array 302, and independently configure a filter group, so that each channel or each group of channels can more freely collect fluorescence in a specific band. Similarly, the LED excitation light can also pass through each optical fiber or each group of imaging fiber arrays to reach the bottom of the cell culture well plate to be excited, and perform fluorescence excitation on cells containing fluorescent dyes.

[0074] In addition, when the temperature and humidity sensor detects that the temperature in the cell operation space is lower than the set temperature threshold, it means that the temperature in the cell operation space is low. First, the flip motor 406 is used to flip the hot end of the thermoelectric cooling plate 405 to face the inside of the insulation frame 404, and the thermoelectric cooling plate 405 is started to heat the heat exchange tube 407. Then, the circulating pump 409 is used to extract the flow medium in the liquid tank 408 and pump it into each heat exchange tube 407 through the hose and the diversion pipe. When the flow medium flows through the insulation frame 404 during the process of flowing in the heat exchange tube 407, it is heated by the heat exchange frame 404. The high temperature environment is heated to a preset temperature threshold, so that the temperature of the flowing medium in the heat exchange tube 407 reaches the preset temperature threshold, and then the flowing medium enters the heat conduction tube 401 through the liquid inlet pipe 402 and exchanges heat with the heat spreader 410. After the temperature of the heat spreader 410 reaches the preset temperature threshold, the interior of the cell operation space is heated, thereby increasing the temperature inside the cell operation space to the set temperature threshold. When the interior of the cell operation space needs to be cooled, the cold end of the thermoelectric cooling plate 405 is directed toward the interior of the insulation frame 404 and operated in the above manner.

[0075] In addition, when the gas sensor detects that the content of a certain gas (such as oxygen) in the cell operation space is low, oxygen needs to be added to the cell operation space. At this time, the solenoid valve on the top of the gas compression tank containing oxygen is opened, allowing the oxygen to pass through the gas collecting pipe 502 into the gas guide pipe 503, and then enter the introduction pipe 508 located on one side, and be sprayed into the cell operation space through the nozzle 509 to achieve the purpose of gas supply. At this time, the nozzle 509 on the introduction pipe 508 is an air nozzle.

[0076] When the temperature and humidity sensor detects that the humidity in the cell operation space is lower than the set threshold, the water pump 505 is started to extract the liquid in the water tank 504, and after being pressurized through the guide pipe 506, it is pumped into the inlet pipe 508 on the other side. When the liquid is sprayed out through the nozzle 509, since the nozzle 509 here is an atomizing nozzle, the liquid enters the cell operation space in an atomized state to achieve the purpose of humidification.

[0077] Example 2

[0078] In actual use, when humidifying and supplying gas to the cell operation space, the humidification in the cell operation space will be uneven, and the injected gas will be unevenly distributed in the cell operation space. Therefore, in order to solve the above problems, the following improvements are made:

[0079] See also Figure 5-8 An adjusting gear 510 is fixedly sleeved on the middle part of the inlet pipe 508 on both sides.

[0080] While humidifying or filling the cell operation space with gas, the rotating motor 214 is started to drive the slide rail 205 to rotate 90 degrees through the electric telescopic rod 203 and the frame seat 204, so that the slide rail 205 and the horizontal electric slide rail 202 are in a vertical state in the horizontal direction. Then, the driving motor 213 is started to drive the slide rail 205 to move horizontally through the engagement of the transmission gear 212 and the tooth plate 210, so that one end of the slide rail 205 moves toward the inlet pipe 508 on one side for humidification or gas supply. Secondly, the electric push rod 507 is started to drive the inlet pipe 508 on this side to move downward, so that the adjustment gear 510 is correspondingly engaged with the tooth plate 210. Then, while humidifying or filling the gas, the driving motor 213 is rotated back and forth, so that the slide rail 205 and the tooth plate 210 reciprocate. The inlet pipe 508 can be rotated back and forth by adjusting the gear 510, and the rotation angle is less than 180 degrees, so that the humidified airflow or the filled gas is evenly distributed in the cell operation space.

[0081] However, during the process of humidifying or adding gas to the cell manipulation space, the temperature of the humidified airflow or added gas differs significantly from the temperature inside the cell manipulation space. This can cause large temperature fluctuations inside the cell manipulation space, affecting the reliability of the experimental results. Therefore, the following improvements are made:

[0082] Arc plates 411 are symmetrically provided on both sides of the heat spreader 410 , and magnetic strips 412 are embedded on the arc sides of the arc plates 411 . The arc plates 411 are made of elastic material and are used to guide the airflow to the vicinity of the plate slot 105 .

[0083] Magnetic blocks are provided at both ends of the slide rail 205 .

[0084] When in use, based on the above operation, after the gear 510 is adjusted to mesh with the tooth plate 210, the drive motor 213 is used to drive the transmission gear 212 to drive the tooth plate 210 to move linearly, so that the nozzle 509 rotates 180 degrees. At this time, the nozzle 509 corresponds to the bottom surface of the heat plate 410. The drive motor 213 is then used to drive the slide rail 205 to reciprocate so that the introduction tube 508 drives the nozzle 509 to swing back and forth. Since the temperature of the heat plate 410 is the same as that in the cell operation space, the nozzle 509 rotates 180 degrees. While the nozzle 508 is swinging back and forth, the humidified airflow or gas ejected from the nozzle 509 first contacts the heat spreader 410 and exchanges heat with the heat spreader 410, thereby making the temperature of the humidified airflow or gas ejected from the nozzle 509 the same as or similar to the temperature in the cell operation space. During the reciprocating swinging of the inlet tube 508, the ejected humidified airflow or gas is temperature-compensated and then discharged to the plate slot 105 along the curved plates 411 on both sides of the heat spreader 410, thereby effectively preventing large temperature fluctuations in the cell operation space.

[0085] Furthermore, while the slide rail 205 reciprocates, the magnetic block at the end of the slide rail 205 repeatedly approaches the magnetic strip 412 on the side of the arc plate 411, and then under the action of the magnetic force between the magnetic strip 412 and the magnetic block, the bottom of the arc plate 411 swings back and forth, so that the humidified airflow or gas flowing along the inner side of the arc plate 411 is evenly discharged into the cell operation space, thereby avoiding large temperature fluctuations in the cell operation space while making the humidified airflow and gas evenly distributed.

[0086] It is worth noting that when the humidified airflow or gas contacts the heat spreader 410 for heat exchange, the temperature of the heat spreader 410 will fluctuate, resulting in the humidified airflow or gas contacting the heat spreader 410 unable to meet the experimental temperature requirements. Therefore, when the humidified airflow or gas contacts the heat spreader 410, the power of the thermoelectric cooling plate 405 is increased to pre-compensate for the temperature fluctuation of the heat spreader 410.

[0087] Example 3

[0088] In addition, when the humidity in the cell operation space is too high, it will interfere with the experiment and reduce the reliability of the experimental results. Therefore, the following improvements are made:

[0089] One of the gas compression tanks in the gas supply tank group 501 is pre-filled with relatively dry air that meets the humidity requirements of the cell operation space.

[0090] The water pump 505 is preferably a bidirectional water pump, that is, the water pump 505 can work in both forward and reverse directions.

[0091] When the temperature and humidity sensor detects that the humidity in the cell operation space is too high, the solenoid valve corresponding to the gas compression tank filled with relatively dry air is opened, and the dry air is released and enters the inlet pipe 508 for gas injection on one side through the air guide pipe 503. At the same time, since dry air will sink and high-humidity air will float in a closed space, during the process of injecting dry air into the cell operation space, the electric push rod 507 corresponding to the inlet pipe 508 through which dry air circulates is started to extend, so that the inlet pipe 508 moves downward to a position close to the operating table 101, and at this time the inlet pipe 508 for humidification is still located at the top of the cell operation space, thereby The dry air discharged into the cell operation space is firstly made to reach the bottom of the cell operation space. With the continuous discharge of dry air, the high-humidity air can be squeezed upwards. At the same time, the water pump 505 is started and made to work in reverse, so that the inlet pipe 508 for top humidification and the nozzle 509 on the inlet pipe 508 are in a negative pressure state. The high-humidity gas gathered at the top of the cell operation space can be sucked into the inlet pipe 508 through the nozzle 509 and reversely input into the water tank 504 through the port of the water pump 505, thereby completing the dehumidification of the interior of the cell operation space. The top of the water tank 504 is connected to the outside world, and the airflow discharged into the water tank 504 can be discharged.

[0092] Example 4

[0093] The mapping system also includes a cell suspension module, which uses a peristaltic pump and a multi-way selection valve to load cells, replenish liquid, recover waste liquid, and recover overflow liquid in the cell container in the cell operation space.

[0094] Each port of the multi-way selection valve is connected to a container. After the suspended cells and different solutions are pumped into the multi-way selection valve by a peristaltic pump, the mixed cell solution is finally pumped into the cell container in the cell operation space.

[0095] The cell operation space is equipped with an independent recovery tank for collecting waste liquid and overflow. The waste liquid generated during the experiment is pumped out of the cell operation space via a peristaltic pump and finally reaches the waste liquid recovery container through a multi-way selection valve. The overflow generated during the experiment flows directly back to the overflow recovery container through the overflow tank of the cell container.

[0096] The above descriptions are merely embodiments of the present invention and are not intended to limit the patent scope of the present invention. Any equivalent structure or equivalent process transformation made using the contents of the present invention description and drawings, or directly or indirectly applied in other related technical fields, are also included in the patent protection scope of the present invention.

Claims

1. A high-throughput fluorescence mapping system based on an imaging fiber array, characterized in that: The system includes a box, a clamping module, an imaging module, and a cell culture well plate. An operating table is provided in the middle of the inner side of the box, which divides the inner space of the box into two parts: the upper part of the operating table is the cell operation space, and the lower part of the operating table is the cell imaging space. A plate slot is provided on the top of the operating table; the imaging module is located in the cell imaging space. The clamping module is arranged in the cell operation space, and the clamping module includes a longitudinal electric slide rail and a transverse electric slide rail. There are two longitudinal electric slide rails, which are respectively arranged on both sides of the inner top surface of the box body. The top ends of the transverse electric slide rail are respectively fixedly connected to the sliding parts of the longitudinal electric slide rails on both sides. The bottom of the sliding part of the transverse electric slide rail is provided with an electric telescopic rod, and the bottom end of the electric telescopic rod is provided with a frame seat. The bottom of the frame seat is slidably connected to the slide rail, and the internal rotation of the slide rail is connected to a bidirectional screw. Both sides of the bidirectional screw are threaded with clamping plates; the bottom of the sliding part of the transverse electric slide rail is embedded with a rotating motor, and the output shaft of the rotating motor is fixedly connected to the fixed end of the electric telescopic rod; both ends of the slide rail are provided with magnetic blocks; A temperature control module is provided on the top of the box, which includes a heat pipe, a heat spreader is provided on the bottom of the heat pipe, and curved plates are symmetrically provided on both sides of the heat spreader, and magnetic strips are embedded on the curved sides of the curved plates; A gas regulating module is provided on the top of the box body, which includes a gas supply tank group, a water tank and an electric push rod. The water tank is provided on the top surface of the box body, and a water pump is provided on one side of the water tank; the electric push rods are symmetrically provided in two groups, each group contains two, and the two groups of electric push rods are symmetrically provided on both sides of the top surface of the box body. An inlet pipe is provided between the bottom telescopic ends of the two electric push rods in the same group, and nozzles are evenly provided on the inlet pipe; the gas supply tank group includes several kinds of gas compression tanks, and the tops of the various gas compression tanks in the gas supply tank group are connected together through a solenoid valve. It is connected to an air collecting pipe, on which an air guide pipe is provided. The input port of the water pump is fixedly connected to the water tank, and the output port of the water pump is provided with a flow guide pipe. One end of the inlet pipes on both sides is fixedly connected to the air guide pipe and the flow guide pipe respectively, and the middle of the inlet pipes on both sides is fixedly sleeved with an adjusting gear; the adjusting gear is used to engage with the tooth plate to rotate the nozzle to correspond to the bottom surface of the heat spreader, and the driving motor makes the inlet pipe swing back and forth. When the inlet pipe swings back and forth, the humidified airflow or gas ejected flows along the arc plate after temperature compensation and is discharged to the plate position slot.

2. The high-throughput fluorescence mapping system based on an imaging fiber array according to claim 1, characterized in that: The imaging module includes a sealing cover, an imaging fiber optic array, a focusing module, a spectrometer module, a light source module, an imaging camera and an imaging fiber optic array lens; the imaging fiber optic array is an array composed of several imaging optical fibers, one end of the imaging fiber optic array is gathered and the other end is dispersed, the imaging fiber optic array lens is a rectangular array arranged at the bottom of the middle plate slot, the sealing cover is arranged at the bottom of the operating table and at a position corresponding to the middle plate slot, the top of the imaging fiber optic array passes through the bottom of the sealing cover, the spectrometer module is arranged at the bottom end of the imaging fiber optic array, the light source module is arranged on one side of the spectrometer module, and the imaging camera is arranged on the outer surface of the spectrometer module; the outside of the imaging fiber optic array is fixedly sleeved with a limiting cylinder, the outside of the limiting cylinder is provided with a support, and the support is arranged on the inner bottom surface of the box; the imaging fiber optic array lens corresponds one-to-one to the bottom of the hole in the cell culture well plate, the optical fiber at one end of the imaging fiber optic array located inside the sealing cover is divergent and connected to the bottom of each imaging fiber optic array lens, and the focusing module is arranged between the imaging fiber optic array and the spectrometer module.

3. The high-throughput fluorescence mapping system based on an imaging fiber array according to claim 2, characterized in that: The spectrometer module is equipped with a dichroic mirror and filters to separate light of different wavelengths. The imaging camera is a high-speed camera with a shooting rate greater than 400 FPS to capture changes in high-speed fluorescence signals. Several imaging cameras are installed outside the spectrometer module to simultaneously record and collect various fluorescence images.

4. The high-throughput fluorescence mapping system based on an imaging fiber array according to claim 1, characterized in that: The two sides of the bidirectional screw are symmetrically provided with threads with opposite spiral directions. The top of the clamping plate is slidably matched with the slide rail. A clamping motor is provided at one end of the slide rail. The output shaft of the clamping motor is fixedly connected to the end of the bidirectional screw. The bottom ends of the clamping plates on both sides are symmetrically provided with clamping seats, and arc grooves are opened on the opposite sides of the clamping seats on both sides.

5. The high-throughput fluorescence mapping system based on an imaging fiber array according to claim 1, characterized in that: A tooth plate is provided on the top of the slide rail, and a transmission gear is connected to the inside of the frame seat through a rotating shaft. The transmission gear is engaged with the tooth plate. A drive motor is provided on the side of the frame seat, and the output shaft of the drive motor is fixedly connected to the end of the rotating shaft.

6. The high-throughput fluorescence mapping system based on an imaging fiber array according to claim 1, characterized in that: A side groove is provided on the side of the box at the position corresponding to the cell operation space. A door panel is movably provided on one side of the side groove, and a temperature and humidity sensor and a gas sensor are installed on the door panel. One side of the box is transparent, and a sealed door is movably provided on the transparent side of the box. There are five plate slots, and the bottom of the plate slot in the middle is a hollow structure, and cell culture well plates are placed in the plate slots.

7. The high-throughput fluorescence mapping system based on an imaging fiber array according to claim 1, characterized in that: The temperature control module also includes an insulation frame and a liquid tank. The heat conduction pipe is arranged on the internal top side of the box body, and the two ends of the heat conduction pipe are respectively provided with a liquid inlet pipe and a liquid return pipe that pass through the top surface of the box body; the insulation frame is arranged on the top surface of the box body, and the internal top side of the insulation frame is rotatably connected to a thermoelectric cooling plate, and the bottom of the insulation frame is horizontally penetrated by a heat exchange tube in a linear array, and the two ends of the heat exchange tube are respectively provided with a collecting pipe and a shunt pipe; the liquid tank is arranged on the top surface of the box body, and a circulating pump is provided on the liquid tank.

8. The high-throughput fluorescence mapping system based on an imaging fiber array according to claim 7, characterized in that: The heat conduction pipe is arranged in an S shape, the arc plate is made of elastic material, a flip motor is provided on the side of the insulation frame, and the output shaft of the flip motor is fixedly connected to the middle of the end face of the thermoelectric cooling plate; the collecting pipe is fixedly connected to the top end of the liquid inlet pipe; the input port of the circulating pump is fixedly connected to the liquid tank, and the output port of the circulating pump is fixedly connected to the diversion pipe through a hose.

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