High-throughput living cell imager for customizing cell physiological environment

Abstract

The utility model discloses a high-flux living cell imager for customizing a cell physiological environment. Relates to the technical field of living cell analysis. The XY electric displacement platform and the automatic focusing module are arranged below the closed cell culture chamber; the multi-color fluorescent module is arranged in the automatic focusing module and comprises a fluorescent bottom shell, single-channel fluorescent assemblies, an adjusting assembly and a fluorescent stepping motor, a camera is arranged below the fluorescent bottom shell, the single-channel fluorescent assemblies are different in color, the adjusting assembly is connected with the single-channel fluorescent assemblies, and the fluorescent stepping motor is arranged at the bottom of the fluorescent bottom shell. According to the utility model, specimens are placed in the closed cell culture chamber; the X-Y electric displacement platform moves to the position below a specimen, focusing is achieved through the automatic focusing module, the adjusting assembly is matched with the fluorescence stepping motor to adjust the single-channel fluorescence assemblies of different colors, and fluorescence images of different colors are shot.

Description

Technical Field

[0001] The utility model relates to the field of live cell analysis, and more specifically to a high-throughput live cell imager for customizing the cell physiological environment. Background Art

[0002] The live cell detection technology refers to the technology of using advanced biological, biochemical, biophysical and other technical means to monitor and analyze live somatic cells in real time and dynamically. This technology has a wide range of applications in the fields of life science, medical research, drug development and so on.

[0003] Traditional cell detection techniques usually analyze cells at fixed time points, unable to achieve real-time dynamic monitoring of cells. Therefore, it is difficult to capture the dynamic changes and reaction processes of cells, and key points of cell activities will also be missed. With the in-depth development of biological and medical research, scientists' demand for real-time monitoring and analysis of biological processes inside living cells has been increasing day by day, which has promoted the development of living cell detection techniques. Current living cell detection techniques include: ① Optical imaging techniques: including fluorescence microscopes, confocal microscopes, two-photon microscopes, etc.; ② Flow cytometry techniques: By suspending single cells in a fluid and using means such as lasers or electric fields to detect and analyze cells, high-throughput cell analysis can be achieved; ③ Fluorescent probe techniques: By designing and synthesizing fluorescent probes with specific biological activities, real-time monitoring of various biomolecules inside cells can be achieved, such as ion concentration, molecular signal transduction, etc.; ④ Microfluidic techniques: Using microfluidic chips to precisely manipulate and analyze cells, achieving control and regulation of the cell microenvironment, as well as evaluation of cell functions. Currently, living cell detection techniques are still constantly developing and improving, providing powerful tools and technical support for fields such as life science research, medical diagnosis, and drug development. However, none of the aforementioned detection techniques can detect the entire process, for a long time, and without interference of the changes in living cells, nor can they be detected in a specific physiological environment, let alone provide a perfect control and detection system. With the development of life science, it has now been clarified that different cells require different physiological environments, especially the oxygen concentration. For example, the physiological oxygen concentration of lung cells is 5.6%, that of kidney cells is 9.5%, and that of large intestine cells is 7.6%. Recently, the research hotspots in the field of cells are to try to restore the original physiological environment of cells or artificially create specific physiological environments, such as the physiological environment of high-altitude hypoxia and cell ischemia-reperfusion, and study the change processes of living cells in this environment. Currently, other solutions basically rely on a three-gas cell culture incubator to maintain the cell state. When detection is needed, the cells are taken out of the incubator for microscopic detection, flow cytometry detection, or placed on other instruments for detection. In this way, the cells lose their previous state, resulting in experimental errors and being prone to cell contamination. Even so, the detected data are only data at certain time points, rather than the complete and real change process without external interference.

[0004] For example, the adverse effects of ischemic tissue damage and similar diseases on the functional status of patients are becoming increasingly severe, and have become the main growing cause of disability and death globally. A key method for studying such diseases and finding treatment solutions is to establish a cell model, that is, to artificially control and programatically change the O2 concentration of its environment during cell culture, creating a culture environment of first ischemia / hypoxia and then reoxygenation (professionally known as ischemia-reperfusion), so as to continuously observe and analyze the changes of cells, evaluate their damage, and a control group can be set up for drug intervention, analyze the differences between the experimental group and the control group to screen suitable drugs and suitable treatment procedures. However, currently there is no instrument that can programatically control the physiological environment of cells throughout the process while observing and analyzing various indicators of cells.

[0005] Therefore, how to provide a high-throughput live cell imager for customizing the cell physiological environment that can programatically control the physiological environment of cells throughout the process while observing and analyzing various indicators of cells is an urgent problem to be solved by those skilled in the art. Summary of the Utility Model

[0006] In view of this, the present utility model provides a high-throughput live cell imager for customizing the cell physiological environment, aiming to solve one of the problems in the above background technology, and capable of programatically controlling the physiological environment of cells throughout the process while observing and analyzing various indicators of cells.

[0007] To achieve the above object, the present utility model provides a high-throughput live cell imager for customizing the cell physiological environment, comprising:

[0008] A housing, a sealed cell culture chamber is provided on the top of the housing, and a CO 2 gas supply port and an N 2 gas supply port are provided on the side wall of the housing;

[0009] An XY electric displacement platform and an autofocus module, the XY electric displacement platform and the autofocus module are arranged in the housing, and the XY electric displacement platform and the autofocus module are arranged below the sealed cell culture chamber, and the autofocus module is arranged on the XY electric displacement platform;

[0010] Multi-color fluorescence module, the multi-color fluorescence module is arranged inside the autofocus module, the multi-color fluorescence module includes a fluorescence bottom case, a single-channel fluorescence component, an adjustment component and a fluorescence stepper motor, a camera is arranged below the fluorescence bottom case, a linear guide rail is arranged at the inner bottom of the fluorescence bottom case, a pressing block is arranged on the linear guide rail, there are multiple single-channel fluorescence components, the colors of the single-channel fluorescence components are different, the single-channel fluorescence components are all arranged on the linear guide rail through the pressing block, the adjustment component is arranged inside the fluorescence bottom case, the adjustment component is connected with the single-channel fluorescence component, the fluorescence stepper motor is arranged at the bottom of the fluorescence bottom case, and a fluorescence cover plate is arranged at the top of the fluorescence bottom case.

[0011] Further, the sealed cell culture chamber includes a chamber bottom plate, a chamber cover plate and a drawer bottom plate. The chamber bottom plate is arranged on the top of the machine shell. An air filter, a temperature sensor, a humidity sensor, a CO 2 concentration sensor and an O 2 concentration sensor are arranged at the inner bottom of the chamber bottom plate. A transparent glass is arranged at the bottom of the chamber bottom plate. A linear rail is arranged at the top of the chamber bottom plate. The drawer bottom plate is slidably arranged on the linear rail. Multiple observation vessels are arranged on the drawer bottom plate. The chamber cover plate is arranged on the chamber bottom plate. A buckle is arranged between the chamber cover plate and the drawer bottom plate. An air filter exhaust port, an N 2 intake hole, a CO 2 intake hole, a double heating module and a lamp board are arranged at the top of the chamber cover plate. The air filter is communicated with the air filter exhaust port, and the air filter exhaust port is communicated with the outside of the machine shell. The CO 2 intake hole is communicated with the CO 2 gas supply port. The N 2 intake hole is communicated with the N 2 gas supply port.

[0012] Further, the XY electric displacement platform includes a Y-axis base, an X-axis base and a Z-axis base. First grating scales, first grating reading heads and first limit switches are arranged on both side walls of the Y-axis base. A first stepper motor and a first guide rail are arranged on the Y-axis base. The X-axis base is slidably arranged on the first guide rail. Second grating scales, second grating reading heads and second limit switches are arranged on both side walls of the X-axis base. A second stepper motor and a second guide rail are arranged on the X-axis base. The Z-axis base is slidably arranged on the second guide rail. A placement groove is arranged on the Z-axis base.

[0013] Further, the autofocus module includes a stepper screw motor, a converter fixing plate, and an electric converter. The stepper screw motor is disposed at the bottom of the Z-axis base. The converter fixing plate is slidably disposed in the placement groove. A vertical guide rail is provided between the converter fixing plate and the placement groove. The multi-color fluorescence module is disposed in the converter fixing plate. The electric converter is disposed on the converter fixing plate. The multi-color fluorescence module is located directly below the electric converter. The output end slider of the focusing stepper screw motor is connected to the converter fixing plate.

[0014] Further, a plurality of objective lenses are provided on the electric converter. The magnifications of the plurality of objective lenses are different. The multi-color fluorescence module is located directly below the plurality of objective lenses with different magnifications. A laser autofocus sensor is further provided on the electric converter. The laser autofocus sensor is used to measure the distance to different specimens.

[0015] Further, the adjusting assembly includes a driving synchronous pulley and an idler pulley. Two driving synchronous pulleys are provided. One of the driving synchronous pulleys is connected to the shaft end of the fluorescence stepper motor. A synchronous belt is sleeved between the two driving synchronous pulleys. A plurality of pressing blocks are provided on the synchronous belt. Each single-channel fluorescence assembly is connected to the synchronous belt through the pressing block. An idler pulley shaft is provided at one end of the driving synchronous pulley away from the fluorescence stepper motor. The idler pulley is sleeved on the idler pulley shaft through a fixing member. The fixing member is connected to the fluorescence bottom case through a tension adjusting screw.

[0016] Further, each single-channel fluorescence assembly includes a first housing, a filter, a dichroic mirror, and a second housing. The side wall of the first housing is connected to the pressing block. The filter is cylindrical. The filter is provided at the bottom of the first housing through a setscrew. The dichroic mirror is disposed at one end of the first housing away from the filter. The second housing is disposed at one end of the dichroic mirror away from the first housing. The second housing is connected to the first housing through screws. A monochromatic collimated light source is provided at one end of the second housing away from the dichroic mirror. The monochromatic collimated light sources are of different colors in different single-channel fluorescence assemblies.

[0017] Further, the dichroic mirror is disposed between the first housing and the second housing and inclined at 45° towards the monochromatic collimated light source.

[0018] Further, it further includes an exhaust fan and a control circuit board. The exhaust fan is disposed on the side wall of the machine case. The exhaust fan is communicated with the interior of the machine case. The control circuit board is also disposed on the side wall of the machine case. The control circuit board is electrically connected to the multi-color fluorescence module.

[0019] As can be seen from the above technical solutions, compared with the prior art, the present utility model discloses a high-throughput live cell imager for customizing the cell physiological environment. By placing cells in a sealed cell culture chamber, the carbon dioxide, oxygen concentration and humidity environment in the sealed cell culture chamber are adjusted to provide an environment required for maintaining the growth conditions and physiological activities of live cells or organoids. The autofocus module is moved in the X-axis and Y-axis directions by an XY electric displacement platform to move the autofocus module to the position where shooting is required, and autofocus is performed through the autofocus module. By adjusting the components and the fluorescence stepping motor, multiple single-channel fluorescence components of different colors in the multi-color fluorescence module are quickly switched. When a certain color of fluorescence is required, the single-channel fluorescence component of the corresponding color is switched to directly below the autofocus module to achieve shooting of fluorescence images of different colors. BRIEF DESCRIPTION OF THE DRAWINGS

[0020] In order to more clearly illustrate the technical solutions in the embodiments of the present utility model or the prior art, the following will briefly introduce the drawings required for the description of the embodiments or the prior art. Obviously, the drawings in the following description are only the embodiments of the present utility model. For those of ordinary skill in the art, other drawings can be obtained according to the provided drawings without creative efforts.

[0021] Figure 1 Structural schematic diagram of the high-throughput live cell imager for customizing the cell physiological environment provided by the present utility model;

[0022] Figure 2 Structural schematic diagram of the sealed cell culture chamber provided by the present utility model;

[0023] Figure 3 Exploded view of the sealed cell culture chamber provided by the present utility model;

[0024] Figure 4 Another perspective exploded view of the sealed cell culture chamber provided by the present utility model;

[0025] Figure 5 Internal structural schematic diagram of the sealed cell culture chamber provided by the present invention;

[0026] Figure 6 Structural schematic diagram of the multi-color fluorescence module provided by the present invention;

[0027] Figure 7 Exploded view of the multi-color fluorescence module provided by the present invention;

[0028] Figure 8 Structural schematic diagram of the single-channel fluorescence component provided by the present invention;

[0029] Figure 9Explosion diagram of the single-channel fluorescence component provided by the present invention;

[0030] Figure 10 Schematic structural diagram of the XY electric displacement platform provided by the present invention;

[0031] Figure 11 Schematic structural diagram of the X-axis base and Z-axis base separated from the Y-axis base provided by the present invention;

[0032] Figure 12 Schematic structural diagram of the XY electric displacement platform and the autofocus module provided by the present invention;

[0033] Figure 13 Installation schematic diagram of the XY electric displacement platform provided by the present invention;

[0034] Figure 14 Schematic structural diagram of the autofocus module provided by the present invention;

[0035] Figure 15 Explosion diagram of the autofocus module provided by the present invention.

[0036] Wherein: 1 is the casing; 2 is the sealed cell culture chamber; 3 is the CO 2 gas supply port; 4 is the XY electric displacement platform; 5 is the autofocus module; 6 is the multi-color fluorescence module; 7 is the fluorescence bottom case; 8 is the single-channel fluorescence component; 9 is the fluorescence stepper motor; 10 is the camera; 11 is the linear guide rail; 12 is the pressing block; 13 is the fluorescence cover plate; 14 is the chamber bottom plate; 15 is the chamber cover plate; 16 is the drawer bottom plate; 17 is the air filter; 18 is the temperature sensor; 19 is the humidity sensor; 20 is the CO 2 concentration sensor; 21 is the transparent glass; 22 is the linear guide; 23 is the observation vessel; 24 is the buckle; 25 is the air filter exhaust port; 26 is the CO 2Air inlet hole; 27 is a dual heating module; 28 is a lamp board; 29 is a Y-axis base; 30 is an X-axis base; 31 is a Z-axis base; 32 is a first grating scale; 33 is a first grating reading head; 34 is a first limit switch; 35 is a first stepper motor; 36 is a first guide rail; 37 is a second grating scale; 38 is a second grating reading head; 39 is a second limit switch; 40 is a second stepper motor; 41 is a second guide rail; 42 is a placement groove; 43 is a stepper screw motor; 44 is a converter fixing plate; 45 is an electric converter; 46 is a vertical guide rail; 47 is a screw; 48 is an objective lens; 49 is a laser autofocus sensor; 50 is a driving synchronous pulley; 51 is an idler pulley; 52 is a synchronous belt; 53 is an idler pulley shaft; 54 is a fixing member; 55 is a tension adjustment screw; 56 is a first housing; 57 is a filter; 58 is a dichroic mirror; 59 is a second housing; 60 is a monochromatic collimated light source; 61 is an exhaust fan; 62 is a control circuit board; 63 is a power supply input port; 64 is a heat dissipation vent; 65 is a fixing member; 66 is a solenoid valve; 67 is a mounting plate; 68 is N 2 Gas supply port; 69 is O 2 Concentration sensor; 70 is N 2 Air inlet hole. Detailed implementation mode

[0037] Next, the technical solutions in the embodiments of the present invention will be clearly and completely described in conjunction with the accompanying drawings in the embodiments of the present invention. Obviously, the described embodiments are only a part of the embodiments of the present invention, rather than all the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those of ordinary skill in the art without creative efforts shall fall within the protection scope of the present invention.

[0038] See Figures 1-15 , on the one hand, the embodiments of the present invention disclose a high-throughput live cell imager for customizing the cell physiological environment, including:

[0039] The machine shell 1, a sealed cell culture chamber 2 is arranged at the top of the machine shell 1, and a CO 2 Gas supply port 3 and N 2 Gas supply port 68; Place the cells in the sealed cell culture chamber 2, and provide carbon dioxide for the sealed cell culture chamber 2 through the CO 2 Gas supply port 3, and introduce N 2 into the sealed cell culture chamber 2 through the N 2 Gas supply port 68, and then control the O 2 concentration;

[0040] The XY electric displacement platform 4 and the autofocus module 5 are arranged inside the machine case 1. The XY electric displacement platform 4 and the autofocus module 5 are arranged below the sealed cell culture chamber 2, and the autofocus module 5 is arranged on the XY electric displacement platform 4. The autofocus module 5 is moved in the X-axis and Y-axis directions through the XY electric displacement platform 4 to move the autofocus module 5 to the position where shooting is required, and focusing is performed through the autofocus module 5.

[0041] The multi-color fluorescence module 6 is arranged inside the autofocus module 5. The multi-color fluorescence module 6 includes a fluorescence bottom case 7, a single-channel fluorescence component 8, an adjustment component, and a fluorescence stepping motor 9. A camera 10 is arranged below the fluorescence bottom case 7. A linear guide rail 11 is arranged at the inner bottom of the fluorescence bottom case 7. A pressing block 12 is arranged on the linear guide rail 11. There are multiple single-channel fluorescence components 8, and the colors of the multiple single-channel fluorescence components 8 are different. Each single-channel fluorescence component 8 is arranged on the linear guide rail 11 through the pressing block 12. The adjustment component is arranged inside the fluorescence bottom case 7 and is connected to the single-channel fluorescence component 8. The fluorescence stepping motor 9 is arranged at the bottom of the fluorescence bottom case 7, and a fluorescence cover plate 13 is arranged at the top of the fluorescence bottom case 7. Photographing is performed through the camera 10. The multiple single-channel fluorescence components 8 with different colors inside the multi-color fluorescence module 6 are quickly switched through the adjustment component and the fluorescence stepping motor 9. The movement of the pressing block 12 on the linear guide rail 11 drives the single-channel fluorescence component 8 to move. When a certain color of fluorescence is required, the single-channel fluorescence component 8 of the corresponding color is switched to directly below the autofocus module 5 to realize the shooting of fluorescence imaging of different colors.

[0042] In this embodiment, the sealed cell culture chamber 2 includes a chamber bottom plate 14, a chamber cover plate 15, and a drawer bottom plate 16. The chamber bottom plate 14 is arranged on the top of the machine case 1. An air filter 17, a temperature sensor 18, a humidity sensor 19, an O 2 concentration sensor 69, and a CO 2 concentration sensor 20 are arranged at the inner bottom of the chamber bottom plate 14. A transparent glass 21 is arranged at the bottom of the chamber bottom plate 14. A linear guide rail 22 is arranged at the top of the chamber bottom plate 14. The drawer bottom plate 16 is slidably arranged on the linear guide rail 22. Multiple observation vessels 23 are arranged on the drawer bottom plate 16. The chamber cover plate 15 is arranged on the chamber bottom plate 14. A buckle 24 is arranged between the chamber cover plate 15 and the drawer bottom plate 16. An air filter exhaust port 25, a CO 2 air inlet hole 26, an N 2 air inlet hole 70, a dual heating module 27, and a lamp board 28 are arranged at the top of the chamber cover plate 15. The air filter 17 is communicated with the air filter exhaust port 25, and the air filter exhaust port 25 is communicated with the outside of the machine case 1. The N 2 air inlet hole 70 is communicated with the N 2 air supply port 68, and the CO2 The air inlet hole 26 is connected to the CO 2 supply port 3; the cells are respectively placed on a plurality of observation vessels 23 in the closed cell culture chamber 2, and through the air filter 17, the air filter exhaust port 25, the temperature sensor 18, the humidity sensor 19, the CO 2 inlet hole 26, the CO 2 concentration sensor 20, the O 2 concentration sensor 69 and the dual heating module 27 cooperate with each other to adjust the carbon dioxide, oxygen concentration and humidity environment in the closed cell culture chamber 2, provide the environment required for maintaining the growth conditions and physiological activities of living cells or organoids, or maintain a specific temperature and humidity and a specific gas environment (such as a hypoxic environment simulating the hypoxic reaction of cells on the plateau) in the closed cell culture chamber 2 according to the experimental procedure.

[0043] In this embodiment, the XY electric displacement platform 4 includes a Y-axis base 29, an X-axis base 30 and a Z-axis base 31. First grating scales 32, first grating reading heads 33 and first limit switches 34 are provided on both side walls of the Y-axis base 29. A first stepping motor 35 and a first guide rail 36 are provided on the Y-axis base 29. The X-axis base 30 is slidably arranged on the first guide rail 36. Second grating scales 37, second grating reading heads 38 and second limit switches 39 are provided on both side walls of the X-axis base 30. A second stepping motor 40 and a second guide rail 41 are provided on the X-axis base 30. The Z-axis base 31 is slidably arranged on the second guide rail 41. A placement groove 42 is provided on the Z-axis base 31; the X-axis base 30 is driven to move on the Y-axis base 29 by the first stepping motor 35 and the first guide rail 36; the moving distance of the X-axis base 30 is limited by the second grating scale 32, the second grating reading head 33 and the second limit switch 34; the Z-axis base 31 is driven to move on the X-axis base 30 by the second stepping motor 40 and the second guide rail 41; the moving distance of the Z-axis base 31 is limited by setting the first grating scale 32, the first grating reading head 33 and the first limit switch 34, so that the autofocus module 5 can be moved in the X-axis and Y-axis directions to move the electric converter 45 to directly below the placement specimen observation vessel 23 that needs to be photographed.

[0044] In this embodiment, the autofocus module 5 includes a stepper screw motor 43, a converter fixing plate 44, and an electric converter 45. The stepper screw motor 43 is arranged at the bottom of the Z-axis base 31. The converter fixing plate 44 is slidably arranged in the placement groove 42. A vertical guide rail 46 is arranged between the converter fixing plate 44 and the placement groove 42. A multi-color fluorescence module 6 is arranged in the converter fixing plate 44. The electric converter 45 is arranged on the converter fixing plate 44. The multi-color fluorescence module 6 is located directly below the electric converter 45. A screw rod 47 is arranged between the output end of the stepper screw motor 43 and the electric converter 45. The stepper screw motor 43 drives the screw rod 47 to move vertically, so that the electric converter 45 moves to the working position, that is, directly below the specimen placement observation vessel 23 to be photographed, to complete autofocus.

[0045] In this embodiment, a plurality of objective lenses 48 are arranged on the electric converter 45. The multiples of the plurality of objective lenses 48 are different. The multi-color fluorescence module 6 is located directly below the plurality of objective lenses 48 with different multiples. A laser autofocus sensor 49 is also arranged on the electric converter 45. The laser autofocus sensor 49 is used to measure the distance to different specimens. By turning the laser autofocus sensor 49 on the electric converter 45 to directly below the observation vessel 23 for placing the specimen, the distance from the objective lens 48 to the sample is measured by the laser autofocus sensor 49 to determine whether the objective lens 48 needs to be displaced in the Z-axis direction, and then the electric converter 45 is switched to the objective lens 48 with the required multiple to reach the working position.

[0046] In this embodiment, the adjustment assembly includes a driving synchronous pulley 50 and an idler pulley 51. Two driving synchronous pulleys 50 are arranged. One of the driving synchronous pulleys 50 is connected to the shaft end of the fluorescence stepper motor 9. A synchronous belt 52 is sleeved between the two driving synchronous pulleys 50. A plurality of pressing blocks 12 are arranged on the synchronous belt 52. Each single-channel fluorescence assembly 8 is connected to the synchronous belt 52 through the pressing block 12. An idler pulley shaft 53 is arranged at one end of the driving synchronous pulley 50 away from the fluorescence stepper motor 9. The idler pulley 51 is sleeved on the idler pulley shaft 53 through a fixing member 54. The fixing member 54 is connected to the fluorescence bottom shell 7 through a tension adjustment screw 55. The positions of the plurality of single-channel fluorescence assemblies 8 in the multi-color fluorescence module 6 are adjusted by the idler pulley 51 cooperating with the driving synchronous pulley 50 and the fluorescence stepper motor 9. The fixing member 54 is used to adjust the tightness of the belt.

[0047] In this embodiment, each single-channel fluorescence component 8 includes a first housing 56, a filter 57, a dichroic mirror 58, and a second housing 59. The side wall of the first housing 56 is connected to the pressure block 12. The filter 57 is cylindrical and is arranged at the bottom of the first housing 56 through a setscrew. The dichroic mirror 58 is arranged at one end of the first housing 56 away from the filter 57. The second housing 59 is arranged at one end of the dichroic mirror 58 away from the first housing 56. The second housing 59 is connected to the first housing 56 by screws. A monochromatic collimated light source 60 is arranged at one end of the second housing 59 away from the dichroic mirror 58, and different colors are set for the monochromatic collimated light source 60 in different single-channel fluorescence components 8. The short-wavelength light wave emitted by the monochromatic collimated light source 60 is reflected by the dichroic mirror 58 with a specific wavelength to the specimen. When there is a corresponding fluorescent substance in the specimen, another long-wavelength light will be emitted, and this light wave will pass through the dichroic mirror 58 and the filter 57 to achieve monochromatic fluorescence imaging.

[0048] In this embodiment, the dichroic mirror 58 is inclined at 45° towards the monochromatic collimated light source 60 and is arranged between the first housing 56 and the second housing 59. By arranging the dichroic mirror 58 obliquely, the light emitted by the monochromatic collimated light source 60 can be reflected onto the specimen.

[0049] In this embodiment, it further includes an exhaust fan 61 and a control circuit board 62. The exhaust fan 61 is arranged on the side wall of the casing 1, and the exhaust fan 61 is communicated with the inside of the casing 1. The control circuit board 62 is also arranged on the side wall of the casing 1, and the control circuit board 62 is electrically connected to the multi-color fluorescence module 6. The casing 1 is cooled by the exhaust fan 61, and the automatic imaging is controlled by the control circuit board 62.

[0050] Specific implementation steps:

[0051] Step 1: Place the cells on multiple observation vessels 23 in the sealed cell culture chamber 2 respectively. Through the air filter 17, the air filter exhaust port 25, the temperature sensor 18, the humidity sensor 19, the CO 2 inlet hole 26, the N 2 inlet hole 70, the CO 2 concentration sensor 20, the O 2 concentration sensor 69, and the double heating module 27 cooperate with each other to adjust the carbon dioxide, oxygen concentration and humidity environment in the sealed cell culture chamber 2, and provide the environment required for maintaining the growth conditions and physiological activities of living cells or organoids;

[0052] Step 2: Move the electric converter 45 in the X-axis and Y-axis directions through the first stepping motor and the second stepping motor on the XY electric displacement platform 4, and move the electric converter 45 directly below the observation vessel 23 for placing the specimen that needs to be photographed;

[0053] Step 3: Rotate the laser autofocus sensor 49 on the electric converter 45 to directly below the observation vessel 23 containing the specimen. Measure the distance from the objective lens 48 to the sample through the laser autofocus sensor 49 to determine whether the objective lens 48 needs to be displaced in the Z-axis direction. Then, switch the objective lens 48 of the required magnification on the electric converter 45 to the working position, and drive the lead screw 47 to move vertically to an appropriate position through the stepper lead screw motor 43;

[0054] Step 4: Quickly switch multiple single-channel fluorescence components 8 of different colors in the multi-color fluorescence module 6 through the driving synchronous pulley 50 and the fluorescence stepper motor 9. When a certain color of fluorescence is required, switch the corresponding single-channel fluorescence component 8 to directly below the objective lens 48 to achieve the shooting of fluorescence images of different colors.

[0055] In addition, a power supply input port 63 is provided on the side wall of the housing 1.

[0056] A heat dissipation ventilation port 64 is also provided on the side wall of the housing 1. The heat dissipation ventilation port 64 communicates with the inside of the housing 1, and the exhaust heat fan 61 is arranged in the heat dissipation ventilation port 64 through the fixing member 65.

[0057] CO 2 The gas supply port 3 is connected to the CO 2 gas source. An electromagnetic valve 66 for controlling the CO 2 input is provided on the connecting pipeline between the gas supply port 3 and the CO 2 gas source. 2 The gas supply port 68 is connected to the N

[0058] N 2 gas source. An electromagnetic valve 66 for controlling the N 2 input is provided on the connecting pipeline between the gas supply port 68 and the N 2 gas source. The concentration of O 2 is controlled by inputting N 2 2 O 2 .

[0059] A humidity generator and an oxygen sensor are also provided inside the sealed cell culture chamber 2.

[0060] The oxygen sensor feeds back the oxygen concentration to the control board 62, and the oxygen concentration is adjusted by introducing N 2 .

[0061] The dynamic detection of living cells requires ensuring a suitable growth environment for the cells, including the carbon dioxide concentration, oxygen concentration, temperature inside the sealed cell culture chamber 2, and a certain humidity range; the CO 2 inlet hole 26, CO 2 concentration sensor 20, and N​2 Intake hole 70, O 2 The concentration sensor 69 cooperates with each other. Through the CO 2 feedback of the concentration sensor 20, when the continuously introduced carbon dioxide reaches the preset value, the introduction of CO 2 is stopped. When the continuously introduced nitrogen causes the oxygen to reach the preset value, it stops, ensuring the normal carbon dioxide concentration and oxygen concentration in the sealed cell culture chamber 2. The temperature is controlled by the temperature sensor 18 and the dual heating module 27; the internal humidity of the sealed cell culture chamber 2 is increased by the humidity generator, and the humidity control requirement is to reach the saturated humidity. The parameters are recorded by the humidity sensor 19; when the drawer bottom plate 16 is closed, the gas with impurities in the environment will be brought into the sealed cell culture chamber 2, and the air filter 17 starts to purify the air in the entire sealed cell culture chamber 2, ensuring a clean environment in the sealed cell culture chamber 2. At the same time, CO 2 , N 2 is introduced to promote CO 2 and O 2 to reach the preset concentration value, and the temperature is adjusted synchronously to form a sealed cell culture chamber 2 with constant temperature and humidity and specific CO 2 and O 2 concentration.

[0062] Mounting plates 67 are provided on the inner side walls of both sides of the casing 1, and both ends of the Y-axis base 29 are respectively connected to the two mounting plates 67.

[0063] In this specification, each embodiment is described in a progressive manner. The key point of each embodiment is to illustrate the differences from other embodiments. The same or similar parts among the embodiments can be referred to each other. For the device disclosed in the embodiment, since it corresponds to the method disclosed in the embodiment, the description is relatively simple, and the relevant parts can be referred to the description of the method part.

[0064] The above description of the disclosed embodiments enables those skilled in the art to implement or use the present invention. Various modifications to these embodiments will be obvious to those skilled in the art. The general principles defined herein can be implemented in other embodiments without departing from the spirit or scope of the present invention. Therefore, the present invention will not be limited to these embodiments shown herein, but will be accorded the widest scope consistent with the principles and novel features disclosed herein.

Claims

1. A high-throughput live cell imager for customizing the cell physiological environment, characterized in that: include: A casing, wherein a sealed cell culture chamber is disposed on the top of the casing, and a CO2 gas supply port and a N2 gas supply port are disposed on the side wall of the casing; An XY electric displacement platform and an autofocus module, wherein the XY electric displacement platform and the autofocus module are arranged in the housing, and the XY electric displacement platform and the autofocus module are arranged below the closed cell culture chamber, and the autofocus module is arranged on the XY electric displacement platform; A multi-color fluorescent module, wherein the multi-color fluorescent module is arranged inside the autofocus module, the multi-color fluorescent module comprises a fluorescent bottom shell, a single-channel fluorescent component, an adjustment component and a fluorescent stepping motor, a camera is arranged below the fluorescent bottom shell, a linear guide is arranged at the bottom of the fluorescent bottom shell, a pressure block is arranged on the linear guide, a plurality of single-channel fluorescent components are arranged, the single-channel fluorescent components have different colors, and the single-channel fluorescent components are all arranged on the linear guide through the pressure block, the adjustment component is arranged in the fluorescent bottom shell, the adjustment component is connected to the single-channel fluorescent component, the fluorescent stepping motor is arranged at the bottom of the fluorescent bottom shell, and a fluorescent cover is arranged on the top of the fluorescent bottom shell.

2. The high-throughput live cell imager for customizing cell physiological environment according to claim 1, characterized in that: The closed cell culture chamber comprises a chamber bottom plate, a chamber cover plate and a drawer bottom plate, the chamber bottom plate is arranged on the top of the casing, an air filter, a temperature sensor, a humidity sensor, a CO2 concentration sensor and an O2 concentration sensor are arranged at the bottom of the chamber bottom plate, a transparent glass is arranged at the bottom of the chamber bottom plate, a linear rail is arranged at the top of the chamber bottom plate, the drawer bottom plate is slidably arranged on the linear rail, a plurality of observation vessels are arranged on the drawer bottom plate, the chamber cover plate is arranged on the chamber bottom plate, a buckle is arranged between the chamber cover plate and the drawer bottom plate, an air filter exhaust port, a CO2 air inlet hole, a N2 air inlet hole, a dual heating module and a lamp board are arranged on the top of the chamber cover plate, the air filter is communicated with the air filter exhaust port, the air filter exhaust port is communicated with the outside of the casing, the CO2 air inlet hole is communicated with the CO2 air supply port, and the N2 air inlet hole is communicated with the N2 air supply port.

3. The high-throughput live cell imager for customizing cell physiological environment according to claim 1, characterized in that: The XY electric displacement platform includes a Y-axis base, an X-axis base and a Z-axis base. The two side walls of the Y-axis base are provided with a first grating scale, a first grating reading head and a first limit switch. The Y-axis base is provided with a first stepper motor and a first guide rail. The X-axis base is slidably arranged on the first guide rail. The two side walls of the X-axis base are provided with a second grating scale, a second grating reading head and a second limit switch. The X-axis base is provided with a second stepper motor and a second guide rail. The Z-axis base is slidably arranged on the second guide rail, and a placement groove is provided on the Z-axis base.

4. A high-throughput live cell imager for customizing cell physiological environment according to claim 3, characterized in that: The autofocus module includes a stepper screw motor, a converter fixing plate and an electric converter, the stepper screw motor is arranged at the bottom of the Z-axis base, the converter fixing plate is slidably arranged in the placement groove, a vertical guide rail is arranged between the converter fixing plate and the placement groove, the multi-color fluorescent module is arranged in the converter fixing plate, the electric converter is arranged on the converter fixing plate, the multi-color fluorescent module is located directly below the electric converter, and the output end slider of the stepper screw motor is connected to the converter fixing plate.

5. A high-throughput live cell imager for customizing cell physiological environment according to claim 4, characterized in that: The electric converter is provided with a plurality of objective lenses with different magnifications. The multicolor fluorescence module is located directly below the objective lenses with different magnifications. The electric converter is also provided with a laser autofocus sensor for measuring the distance to different specimens.

6. The high-throughput live cell imager for customizing cell physiological environment according to claim 1, characterized in that: The adjustment component includes an active synchronous pulley and an idler pulley. Two active synchronous pulleys are provided, one of which is connected to the shaft end of the fluorescent stepper motor. A synchronous belt is sleeved between the two active synchronous pulleys. A plurality of pressure blocks are provided on the synchronous belt. Each single-channel fluorescent component is connected to the synchronous belt through the pressure block. An idler shaft is provided at one end of the active synchronous pulley away from the fluorescent stepper motor. The idler is sleeved on the idler shaft through a fixing member, and the fixing member is connected to the fluorescent bottom shell through a tensioning adjustment screw.

7. The high-throughput live cell imager for customizing cell physiological environment according to claim 6, characterized in that: Each of the single-channel fluorescent components includes a first shell, a filter, a dichroic mirror, and a second shell. The side wall of the first shell is connected to the pressing block. The filter is cylindrical. The filter is arranged at the bottom of the first shell through a top screw. The dichroic mirror is arranged at an end of the first shell away from the filter. The second shell is arranged at an end of the dichroic mirror away from the first shell. The second shell is connected to the first shell through screws. A monochromatic collimated light source is arranged at an end of the second shell away from the dichroic mirror. The monochromatic collimated light source is arranged in different colors in different single-channel fluorescent components.

8. The high-throughput live cell imager for customizing cell physiological environment according to claim 7, characterized in that: The dichroic mirror is disposed between the first shell and the second shell and is inclined at 45 degrees toward the monochromatic collimated light source.

9. The high-throughput live cell imager for customizing cell physiological environment according to claim 1, characterized in that: It also includes a heat exhaust fan and a control board. The heat exhaust fan is arranged on the side wall of the casing and is connected to the interior of the casing. The control board is also arranged on the side wall of the casing and is electrically connected to the multi-color fluorescent module.

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