Cell culture and microimaging integrated device and cell culture and imaging method

Abstract

The invention discloses a cell culture and microscopic imaging integrated device which adopts an integrated design and comprises an imaging assembly, a fluorescence excitation assembly, an objective lens assembly, an automatic shading assembly, a two-dimensional displacement platform, a cell culture assembly, a bright field light source assembly and a shell, the imaging assembly, the fluorescence excitation assembly, the objective lens assembly, the automatic shading assembly and the two-dimensional displacement platform are integrated in the shell, the cell culture assembly is arranged on the two-dimensional displacement platform, and the bright field light source assembly is arranged above the shell. The device has the advantages that the bright field light source, the fluorescence excitation module, the imaging assembly and the cell culture assembly are integrated together, the device is complete in overall function, compact in structure, free of multiple sets of systems and convenient and fast to operate, the temperature and gas environment needed by cell culture are provided through the cell culture assembly, and the requirement for long-time real-time observation of living cell imaging is met; cell culture, multichannel fluorescence imaging and analysis can be completely realized through a single device, the operation process is simplified, and the working efficiency is improved.

Description

Technical Field

[0001] The invention specifically relates to a cell culture microscopic imaging integrated device and a cell culture imaging method. Background Art

[0002] Cell biology is an important branch of life sciences. Cell culture and observation are important research methods in cell biology. Live cell imaging is a technology that uses a microscope to observe the dynamic processes and physiological states of living cells. It can provide spatial and temporal information on dynamic events such as intracellular and extracellular signal transduction, cell migration, cell proliferation, and angiogenesis. It is an important research tool in cancer research, drug development, developmental biology and other fields.

[0003] Live cell imaging generally requires long-term real-time observation while ensuring the activity of cells. Therefore, general microscope systems are difficult to meet the needs of live cell imaging.

[0004] Currently, a feasible solution is to place the imaging device in a cell culture incubator to observe while culturing. However, this solution requires the coordination of multiple systems. Cell culture and imaging need to be controlled separately. The device is large in size, making it difficult to miniaturize and integrate. The process is complicated and cumbersome, and the operation is inconvenient. Summary of the invention

[0005] The first technical problem to be solved by the present invention is that the existing live cell imaging field mentioned in the background technology requires the coordination of multiple systems, and cell culture and imaging need to be controlled separately, which makes it difficult to achieve miniaturization and integration of the device.

[0006] The second technical problem to be solved by the present invention is the problem mentioned in the background technology that the existing field of live cell imaging requires the coordination of multiple systems, has a complicated process and is inconvenient to operate.

[0007] In order to solve the above-mentioned first technical problem, a cell culture microscopic imaging integrated device is proposed; it is realized by the following technical scheme: a cell culture microscopic imaging integrated device, comprising a shell, a display screen is arranged on the shell, the display screen controls the operation of the device and displays the cell imaging in the device, and a support frame for installing other components in the device is arranged in the shell; A bright field light source assembly is arranged at the top of the device, and the bright field light source assembly provides illumination for cell microscopic imaging, and the brightness of the bright field light source assembly is adjustable; The cell culture component is arranged in the middle of the device and is located below the bright field light source component. The cells or tissues to be cultured are placed in a cell culture plate in the cell culture component for culture. The culture environment of the cells in the cell culture component is adjustable. The objective lens assembly is arranged on the bottom plate in the housing through a support frame and is located below the cell culture assembly. The objective lens assembly includes an objective lens screw lifting module and an objective lens module. The magnification and clarity of cell observation in the cell culture assembly are adjusted by the objective lens module and the objective lens screw lifting module. An automatic shading component is arranged between the objective lens component and the cell culture component, and the automatic shading component includes two shading plates, and the interval between the two shading plates is adjustable; A fluorescence excitation assembly is arranged below the objective lens assembly, the fluorescence excitation assembly includes a plurality of fluorescence excitation modules and a fluorescence excitation module fixing plate, the fluorescence excitation modules used for fluorescence microscopy imaging are arranged on the fluorescence excitation module fixing plate, and the upper and lower corresponding relationships between different fluorescence excitation modules and the objective lens assembly are adjustable; The imaging component is arranged below the fluorescent excitation component and located at the bottom of the device. The imaging component is connected to the display screen. The imaging component photographs the cells observed in the cell culture component through the objective lens component and transmits the photographed image to the display screen in real time.

[0008] Preferably, the bright field light source assembly includes a bright field light source module and a lifting screw module. The bright field light source module is arranged on the lifting screw module, and the interval between the bright field light source module and the cell culture assembly is adjusted by the lifting screw module. The setting of the lifting screw module facilitates the adjustment of the brightness of the imaging field of view and is easy to use.

[0009] Preferably, the cell culture assembly includes a cell culture plate, a stage, a sealing frame and a sealing cover. The sealing frame is arranged on the upper panel of the shell, the stage is connected to the two-dimensional movable platform, the cell culture plate is placed on the stage through the sealing frame, and the sealing cover is removable to close the sealing frame. Such an arrangement increases the sealing property of the cell culture plate, facilitates providing a suitable culture environment for cell culture, and ensures the culture effect.

[0010] Preferably, a heating film and a temperature sensor are provided on the stage, and the heating film provides a specified culture temperature for the cells in the cell culture component. A gas nozzle joint is provided on the sealing frame, and the gas nozzle joint is connected to the external air pipe to provide a specified gas environment for the cells in the cell culture component. Such a setting is convenient for providing a suitable culture environment for cell culture and ensuring the culture effect.

[0011] Preferably, the two-dimensional mobile platform comprises a transverse moving component, a longitudinal moving component and a mobile platform mounting plate. The transverse moving component and the longitudinal moving component are mounted on the support frame through the mobile platform mounting plate. The stage in the cell culture component is arranged on the longitudinal moving component. The position of the stage is controllably adjustable through the two-dimensional mobile platform. The arrangement of the two-dimensional mobile platform facilitates adjustment of the position of the cell culture plate and the field of view of the microscopic imaging, and is easy to use.

[0012] Preferably, the technical solution of the present invention is that the automatic shading component includes a front support plate and a rear support plate, the front support plate and the rear support plate are arranged on the support frame, and slide grooves are arranged on the front support plate and the rear support plate, and two shading plates are inserted in the slide grooves and distributed opposite to each other, and a movable component is arranged on the shading plate, and the shading plate can slide controllably along the slide groove through the movable component. The setting of the automatic shading component is convenient for adjusting the brightness of the imaging field of view and is easy to use.

[0013] Preferably, the objective lens module includes an objective lens turntable and an objective lens turntable base, a plurality of objective lenses of different magnifications are arranged on the objective lens turntable, the objective lens turntable is arranged on the objective lens turntable base, the objective lens turntable is controllably rotatable relative to the objective lens turntable base by the drive of a turntable drive motor and a gear set, so as to complete the switching of objective lenses of different magnifications; a light transmission hole is arranged on the objective lens turntable base, and the correspondence between the plurality of objective lenses and the light transmission hole is adjusted by the turntable drive motor. Such an arrangement facilitates the adjustment of the magnification of the imaging by changing the objective lenses of different magnifications, and is easy to use.

[0014] Preferably, there are multiple fluorescence excitation modules, and the wavelengths, excitation light filters and emission light filters of the multiple fluorescence excitation modules are different. Such an arrangement facilitates multi-channel fluorescence imaging through multiple different fluorescence excitation modules.

[0015] In a preferred embodiment of the technical solution of the present invention, the imaging component includes a camera, a horizontal optical path pipe, a right-angle optical path block and an imaging lens. One end of the horizontal optical path pipe is connected to the camera, and the other end is connected to the right-angle optical path block. The imaging lens is arranged above the right-angle optical path block. The light from above the imaging lens is received by the CCD of the camera via the imaging lens, the right-angle optical path block and the horizontal optical path pipe. The arrangement of the imaging component facilitates the real-time transmission of the image in the field of view to the display screen, which is convenient for observation and easy to use.

[0016] In order to solve the second technical problem mentioned above, a cell culture imaging method of an integrated cell culture microscopic imaging device is also proposed; the method specifically comprises the following steps: Cell and tissue culture: A1: Place the cells, tissues and culture fluid to be cultured into the cell culture chamber of the cell culture plate, and place them in multiple cell culture chambers for comparison and reference; A2: Place the cell culture plate on the stage and cover it with a sealing cover to seal the cell culture plate; A3: Open the air nozzle joint to exhaust the air in the cell culture plate, and inject the specified air into the cell culture plate. Turn on the heating film and set it to 37°C for heating to provide the cells in the cell culture assembly with a suitable gas environment and temperature environment, and then carry out cell and tissue culture. Live bright-field microscopy: B1: Adjust the transmission mechanism in the fluorescence excitation assembly to translate the fluorescence excitation module as a whole out of the main optical axis. At this time, the positions of the fluorescence excitation module and the objective lens module do not correspond. B2: Turn on the bright field light source module and camera switch; B3: Adjust the distance between the two shading plates in the automatic shading assembly to expose the cell culture plate. At this time, the imaging status can be viewed in real time on the display screen; B4: Adjust the intensity of the bright field light source module through the lifting screw module to adjust the brightness of the field of view, and then use the two-dimensional mobile platform to adjust the position of the cell culture plate and switch the observation field of view; B5: By adjusting the objective lens screw lifting module, the height of the objective lens module in the vertical direction can be adjusted to achieve focusing at different heights. The turntable drive motor controls the rotation of the objective lens turntable to switch different objective lenses to achieve observations with different magnifications. Real-time fluorescence microscopy: C1: Adjust the transmission mechanism in the fluorescence excitation assembly to translate the fluorescence excitation module as a whole to the main optical axis. At this time, the fluorescence excitation module corresponds to the position of the objective lens module. C2: Turn on the bright field light source module and camera switch; C3: Adjust the distance between the two shading plates in the automatic shading assembly to expose the cell culture plate. At this time, the imaging status can be viewed in real time on the display screen; C4: Adjust the intensity of the bright field light source module through the lifting screw module to adjust the brightness of the field of view, and then use the two-dimensional mobile platform to adjust the position of the cell culture plate and switch the observation field of view; C5: By adjusting the objective lens screw lifting module, the height of the objective lens module in the vertical direction can be adjusted to achieve focusing at different heights. The turntable drive motor controls the rotation of the objective lens turntable to switch different objective lenses to achieve observations with different magnifications. C6: Adjust the transmission mechanism in the fluorescence excitation component to move the fluorescence excitation modules containing different wavelengths to the main optical axis to achieve multi-channel fluorescence imaging.

[0017] Compared with the prior art, the present invention has the following beneficial effects: The technical solution of the present invention integrates a bright field light source, a multi-wavelength fluorescence excitation module, an imaging component and a cell culture component. The overall function of the device is complete and the structure is compact. It is convenient and quick to operate without the need for the coordination of multiple systems. The cell culture component can provide the temperature and gas environment required for cell culture to meet the needs of long-term real-time observation of living cell imaging. In addition, the device is also provided with a display screen, and no external computer or other equipment is required. Cell culture, multi-channel fluorescence imaging and analysis can be fully realized through a single device, which simplifies the operation process, reduces the difficulty of operation, and improves work efficiency. BRIEF DESCRIPTION OF THE DRAWINGS

[0018] Figure 1 It is a three-dimensional schematic diagram of the present invention; Figure 2 It is the internal structure diagram of the present invention; Figure 3 It is an exploded view of the internal structure of the present invention; Figure 4 It is a schematic diagram of the optical path of the present invention; Figure 5 is a three-dimensional schematic diagram of an imaging component; Figure 6 It is a three-dimensional schematic diagram of the fluorescence excitation component; Figure 7 This is the exploded diagram of the fluorescence excitation module; Figure 8 is a three-dimensional schematic diagram of an objective lens assembly; Fig. 9 This is an exploded view of the objective lens assembly; Fig.10 It is a three-dimensional schematic diagram of an automatic shading component; Fig.11 It is a three-dimensional schematic diagram of a two-dimensional mobile platform; Fig.12 Exploded view of the cell culture components; Fig.13 It is a three-dimensional schematic diagram of a bright field light source assembly; Description of the accompanying drawings: 1-housing, 11-upper panel, 12-side panel, 13-bottom plate, 14-functional hole, 15-display screen, 16-switch, 17-USB interface, 2-support frame, 21-right support frame, 22-left support frame, 23-front support frame, 3-bright field light source assembly, 31-bright field light source module, 32-lifting screw rod module, 33-support frame, 34-support back plate, 35-screw rod module housing, 4-cell culture assembly, 41-cell culture plate, 42-stage, 43-cell culture chamber, 44-seal Sealing frame, 45-sealing cover, 46-air nozzle joint, 47-sealing frame groove, 48-sealing cover protrusion, 49-stage sealing ring, 5-two-dimensional mobile platform, 51-lateral moving component, 52-longitudinal moving component, 53-connecting plate, 54-screw motor module, 55-first linear slide pair, 56-mobile platform mounting plate, 6-automatic shading component, 61-shading plate, 62-front support plate, 63-rear support plate, 64-rack, 65-slide, 66-motor mounting frame, 7-objective lens assembly, 701-objective lens screw lift Drop module, 702-objective lens module, 703-support frame, 704-mounting frame, 705-objective lens turntable, 706-objective lens turntable base, 707-objective lens mounting hole, 708-positioning plate, 709-photoelectric switch, 710-light transmission hole, 711-turntable drive motor, 712-connecting shaft, 713-objective lens, 8-fluorescence excitation component, 801-fluorescence excitation module, 802-fluorescence excitation module fixing plate, 803-second linear slide pair, 804-first fixed block, 805-synchronous wheel, 806-second fixed block, 807-synchronous belt, 808-synchronous belt fixing plate, 809-control circuit board, 810-control board mounting plate, 811-base housing, 812-fluorescence excitation light source, 813-housing cover, 814-lens, 815-lens mounting ring, 816-excitation light filter, 817-corner light path channel, 818-dichroic mirror, 819-emission light filter, 9-imaging assembly, 91-camera, 92-horizontal light path pipeline, 93-camera fixing frame, 94-light path fixing frame, 95-right-angle light path block, 96-imaging lens. DETAILED DESCRIPTION

[0019] The following will be combined with the attached embodiment of the present invention Figure 1-13 , the technical solutions in the embodiments of the present invention are described in detail. Example

[0020] like Figure 1 , 2As shown in Figures 3 and 4, a cell culture microscopy imaging integrated device is adopted. The device adopts an integrated design and mainly includes a shell 1, a bright field light source component 3, a cell culture component 4, a two-dimensional mobile platform 5, an automatic shading component 6, an objective lens component 7, a fluorescence excitation component 8 and an imaging component 9, wherein the cell culture component 4, the two-dimensional mobile platform 5, the automatic shading component 6, the objective lens component 7, the fluorescence excitation component 8 and the imaging component 9 are integrated inside the shell 1 composed of an upper panel 11, a side panel 12 and a bottom panel 13, the bright field light source component 3 is located above the device and connected to the upper panel 11, and in order to facilitate the stability of the fixation, a support frame 2 for support is also provided in the shell 1, and the support frame 2 can increase the overall stability of the device.

[0021] The main function of the bright field light source assembly 3 is to provide sufficient illumination so that the observed object can be clearly imaged in the field of view. The bright field light source assembly 3 is located above the device and connected to the upper panel 11, and there is a light path in the vertical direction of the device that passes through the bright field light source module 31 in the bright field light source assembly 3 and the center of the imaging lens 96. The light is irradiated along this light path, and this light path is named the main optical axis. The various functional components are distributed in sequence from top to bottom along the main optical axis.

[0022] The main function of the cell culture component 4 is to culture cells or tissues. To facilitate the culture of cells or tissues, the temperature and gas environment in the cell culture component 4 are adjustable.

[0023] The main function of the two-dimensional mobile platform 5 is to adjust the horizontal and vertical positions of the cell culture plate 41 in the cell culture component 4, thereby realizing the switching of the observation field, which is convenient to use.

[0024] The function of the automatic shading component 6 is to cooperate with the bright field light source component 3 to adjust the brightness of the observation field to make the observation clearer.

[0025] The objective lens assembly 7 is an important component of the device. The objective lens 713 therein not only plays the role of magnifying the object, but also can focus the light onto the observed object to form a clear image. The objective lens screw lifting module 701 in the objective lens assembly 7 can adjust the distance between the objective lens 713 and the object to be observed (i.e., the working distance), so that the light can be correctly focused on the object, thereby obtaining a clear image.

[0026] The main function of the fluorescence excitation component 8 is to be used for fluorescence microscopy imaging. The fluorescence excitation component 8 emits light of a specific wavelength (excitation light) to the target to be observed. The fluorescent molecules in the object to be observed will emit fluorescence, and then the target will be imaged by collecting and detecting these fluorescence signals.

[0027] The main function of the imaging component 9 is to capture the imaging field of view and transmit the captured image to the display screen 15 on the housing 1 in real time, so that the user can observe in real time and the device is easy to use.

[0028] Regarding the distribution order of various functional components: the various components arranged on the shell 1 are distributed in sequence from top to bottom along the main optical axis, wherein the bright field light source component 3 is located at the top, and the bright field light source component 3 is installed on the upper panel 11 of the shell 1, the two-dimensional mobile platform 5 is installed inside the shell 1, and is located below the bright field light source component 3, the cell culture component 4 is installed on the two-dimensional mobile platform 5, and is located between the two-dimensional mobile platform 5 and the bright field light source component 3, the automatic shading component 6 is arranged below the two-dimensional mobile platform 5, the objective lens component 7 is arranged in the shell 1, and is located below the automatic shading component 6, the fluorescence excitation component 8 is in the shell 1, and is located below the objective lens component 7, the imaging component 9 is arranged at the bottom of the device, and is located below the fluorescence excitation component 8, and the imaging component 9 is installed on the bottom plate 13 of the shell 1, and during operation, the centers of the fluorescence excitation module 801 and the objective lens 713 in the fluorescence excitation component 8 are also located on the main optical axis; at the same time, the two-dimensional displacement platform 5 has an initial position, so that the center of the cell culture plate 41 installed on the stage 42 is also located on the main optical axis.

[0029] Definition: In this embodiment, the integrated cell culture microscopy imaging device is placed as a whole on a work surface. Based on the work plane where the device is placed, the direction from the work plane to the bright field light source assembly 3 at the end of the device is upward, the direction opposite to it is downward, the direction from the outside to the inside of the device is the inner direction, and on the contrary, the direction from the inside of the device to the outside is the outer direction.

[0030] like Figure 1 and 2 As shown, the shell 1 is a rectangular metal box made of metal plates, and the shell 1 includes an upper panel 11, a bottom plate 12 and four side panels 12. The four side panels 12 form a rectangular frame. The upper panel 11 and the bottom plate 12 use screws to close the rectangular frame to form a hollow shell 1. In order to facilitate real-time display of imaging, a side panel 12 at the front end of the device is tilted, and an installation opening for installing a display screen 15 is opened on this side panel 12. The display screen 15 is installed in this installation opening. The imaging field of view can be displayed in real time through the display screen 15, which is convenient to use. In order to facilitate the operation of the display screen 15, a touch display screen is preferably used.

[0031] In addition, in order to facilitate the switching of the device and the transmission of data, a switch 16 and a USB interface 17 are installed on a side panel 12 on one side of the housing 1 .

[0032] In order to facilitate the placement of the cell culture plate 41 in the cell culture component 4 onto the stage 42 fixed on the two-dimensional movable platform 5, a rectangular functional hole 14 is opened on the upper panel 11 perpendicular to the upper panel 11, and the cell culture plate 41 is placed on the stage 42 through the functional hole 14.

[0033] In order to improve the stability of each component installed in the shell 1, a support frame 2 is also fixed in the shell 1, and the support frame 2 includes a right support frame 21, a left support frame 22 and a front support frame 23. The right support frame 21, the left support frame 22 and the front support frame 23 are all made of metal, wherein the right support frame 21 and the left support frame 22 are the same, the right support frame 21 and the left support frame 22 are connected to the side plate 12 and the bottom plate 13 by screws, the right support frame 21 and the left support frame 22 are distributed on the left and right, and the support frame 23 is connected to the front end of the right support frame 21 and the left support frame 22, and the right support frame 21, the left support frame 22 and the front support frame 23 as a whole form a "U"-shaped frame connected to the shell 1.

[0034] like Figure 1 , 3 As shown in Figure 5, the imaging component 9 includes a camera 91, a horizontal optical path pipe 92, a camera fixing frame 93, an optical path fixing frame 94, a right-angle optical path block 95 and an imaging lens 96, wherein one end of the horizontal optical path pipe 92 is connected to the camera 91, and the other end is connected to one end of the right-angle optical path block 95, and the imaging lens 96 is arranged at the other end of the right-angle optical path block 95. The function of the horizontal optical path pipe 92 and the right-angle optical path block 95 is to transmit light.

[0035] The camera fixing frame 93 is an "L"-shaped metal frame. The camera 91 is installed on the bottom plate 13 by screws through the camera fixing frame 93. The horizontal optical path pipe 92 horizontally connects the camera 91 and the right-angle optical path block 95. There is a "U"-shaped optical path fixing frame 94 below the horizontal optical path pipe 92 to fix and support the horizontal optical path pipe 92. The upper end of the optical path fixing frame 94 is clamped on the horizontal optical path pipe 92, and the lower end is fixed to the bottom plate 13 by screws. A reflector or a reflecting prism is installed at 45° inside the right-angle optical path block 95. An imaging lens 96 is installed above the right-angle optical path block 95. The light from above the imaging lens 96 is focused by the imaging lens 96 and enters the right-angle optical path block 95. After being reflected by the reflector inside the right-angle optical path block 95, it enters the horizontal optical path pipe 92 and is finally reflected by the CCD (charge coupled device (charge coupled device (charge coupled device (charge coupled device (charge coupled device (charge coupled device (charge coupled device (charge coupled device (charge coupled device (charge coupled device (charge coupled device (charge coupled device (charge coupled device (charge coupled device (charge coupled device (charge coupled device (charge coupled device (charge coupled device (charge coupled device (charge coupled device ( The camera 91 is connected to the display screen 15 via a signal line to facilitate real-time observation. The camera 91 transmits the photos taken in the observation field to the display screen 15 in real time.

[0036] like Figure 3 , 6As shown in 7, the fluorescence excitation component 8 includes a fluorescence excitation module 801 and a fluorescence excitation module fixing plate 802. The fluorescence excitation module fixing plate 802 is fixed to the right supporting frame 21 and the left supporting frame 22 by screws. The fluorescence excitation module 801 is arranged on the fluorescence excitation module fixing plate 802, and the position of the fluorescence excitation module 801 on the fluorescence excitation module fixing plate 802 is controllably adjustable.

[0037] The fluorescence excitation module fixing plate 802 includes a first fixing block 804, a second fixing block 806, a control board mounting plate 810 and two mounting plates, one of which is in a long strip shape and the other is in a "U" shape. The two mounting plates are spaced apart, and the two ends of the mounting plates are respectively placed on the beams under the right support frame 21 and the left support frame 22, and then fixed with screws.

[0038] There are multiple groups of fluorescence excitation modules 801, and four groups are preferably used in this embodiment. The wavelengths, excitation light filters 816 and emission light filters 819 of the four groups of fluorescence excitation modules 801 are different. The four groups of fluorescence excitation modules 801 are arranged horizontally, and the ends of the four groups of fluorescence excitation modules 801 are respectively connected by the first fixed block 804 and the second fixed block 806, and the first fixed block 804 and the second fixed block 806 are respectively connected to the mounting plate.

[0039] The control board mounting plate 810 is a "U"-shaped metal plate with a certain thickness. The control board mounting plate 810 is stuck on the base end of the four groups of fluorescence excitation modules 801, and the control board mounting plate 810 is connected to the second fixed block 806. In order to facilitate the control of the fluorescence excitation module 801, a control circuit board 809 is also fixed on the outer side of the control board mounting plate 810, and the opening and adjustment of the fluorescence excitation module 801 are controlled by the control circuit board 809.

[0040] In order to realize the switching between fluorescent field microscopy imaging and bright field microscopy imaging, a position adjustment component is also arranged between the fluorescence excitation module 801 and the fluorescence excitation module fixing plate 802, and the position adjustment component includes a synchronous wheel 805, a second linear slide rail pair 803 and a synchronous belt 807 installed on the synchronous wheel 805, the synchronous wheel 805 has two sets of synchronous wheels respectively installed at both ends of the "U"-shaped mounting plate, and one of the synchronous wheels 805 is connected to a synchronous motor, the second linear slide rail pair 803 has two sets of synchronous wheels respectively installed on the two mounting plates, and the slider on the second linear slide rail pair 803 is connected to the first fixed block 804 and the second fixed block. The stepper motor can drive the two synchronous wheels 805 to move synchronously through the synchronous motor energy and the synchronous belt 807. In order to make the fluorescence excitation module 801 move accordingly, a rectangular synchronous belt fixing piece 808 is fixed on the control board mounting plate 810 with screws. The synchronous belt 807 is connected to the control board mounting plate 810 through the synchronous belt fixing piece 808. The stepper motor movement can drive the fluorescence excitation module 801 to move along the second linear slide rail pair 803, so as to adjust the position of the fluorescence excitation module 801, change the overlap of the fluorescence excitation module 801 with the main optical axis, and realize the switching between fluorescent field microscopy imaging and bright field microscopy imaging.

[0041] A single fluorescence excitation module 801 includes a base housing 811, a fluorescence excitation light source 812, a housing cover 813, a lens 814, a lens mounting ring 815, an excitation light filter 816, a corner light path channel 817, a dichroic mirror 818 and an emission light filter 819. The fluorescence excitation light source 812 is fixed in a cavity formed by the base housing 811 and the housing cover 813, with the emission surface of the fluorescence excitation light source 812 facing outwards. The lens 814 is fixed in the lens mounting ring 815. The lens mounting ring 815 is provided with a plurality of protruding portions. 15 is connected to the outer shell cover 813 through a threaded opening, and the corner light path channel 817 is divided into two parts, the upper right part and the lower left part, by a 45° cut. The upper right part of the corner light path channel 817 has an opening on the right side and the top, and the right side is connected to the other end of the lens mounting ring 815 through a thread, the excitation light filter 816 is installed in the right opening, the lower left part of the corner light path channel 817 has an opening at the bottom, the emission light filter 819 is installed in the lower opening, and the dichroic mirror 818 is embedded in the cut of the corner light path channel 817.

[0042] The excitation light emitted by the fluorescent excitation light source 812 reaches the lens 814 through the opening of the outer shell cover 813, is focused into parallel light by the lens 814, passes through the excitation light filter 816, enters the upper right part of the corner light path channel 817, shines on the dichroic mirror 818, is totally reflected, and is emitted from the upper opening of the corner light path channel 817. In addition, the light emitted from above is emitted to the corner light path channel 817, enters from the upper opening of the corner light path channel 817, shines on the dichroic mirror 818, is transmitted downward, passes through the emission light filter 819, and is emitted from the lower opening.

[0043] The fluorescent excitation light source 812, the dichroic mirror 818 and the filter are all existing devices and can be used directly.

[0044] like Figure 3 , 8 As shown in FIG. 9 , the objective lens assembly 7 includes an objective lens screw lifting module 701 and an objective lens module 702. The objective lens screw lifting module 701 is installed in the housing 1, and the objective lens module 702 is connected to the objective lens screw lifting module 701. The objective lens module 702 can move up and down along the objective lens screw lifting module 701 under the action of the objective lens screw lifting module 701.

[0045] The objective lens screw lifting module 701 includes a screw motor and a moving block installed on the screw, the upper end of the screw motor is connected to the upper panel 11, and the lower end is connected to the bottom plate 13, the moving block is screwed onto the screw, and two rectangular slides are fixed on both sides of the screw with screws, the upper end of the slide is connected to the upper panel 11, and the lower end is connected to the bottom plate 13, the moving block is located in the gap between the two slides, when the screw motor is started, it can drive the moving block to move up and down along the slide, and then drive the objective lens module 702 connected to the moving block to move up and down, so as to adjust the focal length of the field of view.

[0046] In order to improve the stability of the screw motor, an "L"-shaped support frame 703 is fixed on both sides of the screw motor. The support frame 703 is arranged on both sides of the screw motor, and the upper end of the support frame 703 is connected to the upper panel 11, and the lower end is connected to the bottom plate 13.

[0047] In order to facilitate the connection between the objective lens module 702 and the moving block on the objective lens screw lifting module 701, a mounting frame 704 is fixed on the objective lens module 702 with screws. The mounting frame 704 consists of two connecting strips and a connecting block. One end of the two connecting strips is fixed on the connecting block, and the other end is connected to the objective lens module 702 with screws. When the moving block moves up and down, the objective lens module 702 moves up and down.

[0048] The objective lens module 702 includes an objective lens turntable 705, an objective lens turntable base 706, a plurality of objective lenses 713 with different magnifications, and a rotating mechanism for driving the switching of objective lenses, which is composed of gears, a connecting shaft 712 and a turntable driving motor 711.

[0049] The objective lens turntable 705 is shaped as a part of a sphere, and four circular objective lens mounting holes 707 are evenly arranged on the surface of the objective lens turntable 705 around the central axis perpendicular to the sphere. An internal thread is arranged on the inner wall of the objective lens mounting hole 707, and the objective lens 713 is screwed onto the objective lens mounting hole 707.

[0050] The objective lens turntable base 706 is a circular box with a certain height. The upper end of the objective lens turntable base 706 is open. The objective lens turntable 705 is installed in the objective lens turntable base 706 through a connecting shaft 712. At the opening at the upper end of the objective lens turntable base 706, the objective lens turntable 705 can rotate freely relative to the objective lens turntable base 706 around the connecting shaft 712. In addition, in order to facilitate the smooth flow of the light path, a circular light-transmitting hole 710 that penetrates the objective lens turntable base 706 is also opened on the objective lens turntable base 706.

[0051] In order to facilitate the rotation of the objective lens turntable 705, a turntable drive motor 711 is fixed to the objective lens turntable base 706 with screws. The motor shaft of the turntable drive motor 711 passes through the objective lens turntable base 706 and is located inside the objective lens turntable base 706. A gear is fixed on the motor shaft and a gear is also fixed on the connecting shaft 712. The two gears are meshed, and the turntable drive motor 711 can drive the objective lens turntable 705 to rotate.

[0052] In order to facilitate determination of the rotation position of the objective lens turntable 705 so that the objective lens 713 is always aligned with the light-transmitting hole 710, a photoelectric switch 709 with a U-shaped opening is fixedly installed inside the objective lens turntable base 706, and the number of the photoelectric switches 709 is consistent with the number of the objective lenses 713. At the same time, a plurality of positioning plates 708 are fixedly installed under the objective lens turntable 705, and the number of the positioning plates 708 is consistent with the number of the objective lenses 713.

[0053] Regarding the switching of the objective lens 713, when the objective lens 713 needs to be switched, the turntable drive motor 711 rotates to drive the gear to rotate, and then drives the connecting shaft 712 to rotate, thereby driving the objective turntable 705 to rotate. Driven by the rotating objective turntable 705, the positioning piece 708 and the photoelectric switch 709 are in a relative position such that the positioning piece 708 passes through the U-shaped opening of the photoelectric switch 709, thereby blocking the photoelectric switch 709, thereby realizing the rotational positioning of each objective lens 713, and aligning different objective lenses 713 with the light-transmitting holes 710 on the objective turntable base 706, thereby realizing the switching of the lens 713.

[0054] In order to ensure that the objective lens 713 is in a vertical state when aligned with the light-transmitting hole 710, and is exactly located on the main optical axis and parallel to the main optical axis, the connecting strip on the mounting bracket 704 is tilted upward at a certain angle, and at the same time, the angle between the objective lens 73 and the central axis is consistent with the upward tilted angle of the connecting strip on the mounting bracket 704. In this way, when a certain objective lens 713 is rotated to align with the light-transmitting hole 710, it is in a vertical state, and is exactly located on the main optical axis and parallel to the main optical axis.

[0055] like Figure 3 and 10As shown, the automatic shading assembly 6 includes two shading plates 61, a front support plate 62, a rear support plate 63 and a shading plate moving mechanism, the two ends of the front support plate 62 and the rear support plate 63 are respectively fixed on the right support frame 21 and the left support frame 22, and the front support plate 62 and the rear support plate 63 are spaced apart, the two shading plates 61 are inserted into the front support plate 62 and the rear support plate 63 from both ends of the front support plate 62 and the rear support plate 63, and the interval between the two shading plates 61 is adjusted by the shading plate moving mechanism.

[0056] The sunshade 61 is a rectangular metal sheet. Two sunshades 61 are provided. The two sunshades 61 have the same size. The front support plate 62 is a rectangular metal strip with a certain thickness. Threaded holes are provided at both ends of the front support plate 62. The two ends of the front support plate 62 are respectively fixed to the right support frame 21 and the left support frame 22 by screws. In order to facilitate the installation of the sunshade 61, a groove with a rectangular cross-section is provided on the side wall of the front support plate 62 along the length direction of the front support plate 62. This groove is named a slide groove 65. The slide groove 65 is provided along the length direction of the front support plate 62, and the sunshade 61 can slide along the slide groove 65.

[0057] The rear support plate 63 is substantially the same as the front support plate 62, except that there are protrusions perpendicular to the rear support plate 63 at both ends of the rear support plate 63, and the motor in the shading plate moving mechanism is fixed to the protrusions through a motor mounting frame 66, so that the rear support plate 63 as a whole is in a "U" shape. In addition, in order to facilitate the installation of the shading plate 61, a slide groove 65 is also opened on the side wall of the rear support plate 63 along the length direction of the rear support plate 63, and the two ends of the shading plate 61 are respectively inserted into the slide grooves 65 of the front support plate 62 and the rear support plate 63, and the distance between the two shading plates 61 is adjusted by the shading plate moving mechanism.

[0058] The shading plate moving mechanism includes a rack 64, a motor and a motor mounting frame 66. The motor is fixed to the motor mounting frame 66 by screws. The motor mounting frame 66 is fixed to the rear support plate 63, and a gear is also fixed to the main shaft of the motor.

[0059] To facilitate the installation of the rack 64, a rectangular plate protrudes from one end of the shading plate 61 close to the rear support plate 63, and the rack 64 is fixed on this rectangular plate. The rack 64 fixed on the shading plate 61 is meshed with the gear on the motor. The rotation of the motor can control the two shading plates 61 to slide left and right in the horizontal direction, thereby controlling the opening and closing of the automatic shading assembly 6 to achieve the purpose of adjusting the brightness of the field of view.

[0060] like Figure 3 and 11As shown, the two-dimensional moving platform 5 includes a transverse moving component 51 and a longitudinal moving component 52. The transverse moving component 51 is installed on the right supporting frame 21 and the left supporting frame 22 through a moving platform mounting plate 56. The transverse moving component 51 and the longitudinal moving component 52 are connected by a connecting plate 53, wherein the transverse moving component 51 is located at the bottom and the longitudinal moving component 52 is located at the top. The transverse moving component 51 is arranged transversely and the longitudinal moving component 52 is arranged longitudinally. The transverse moving component 51 can drive the longitudinal moving component 52 to move transversely.

[0061] The connecting plate 53 is a metal plate with a rectangular cross section. A rectangular hole is provided on the surface of the connecting plate 53 and penetrates the connecting plate 53. The size of the rectangular hole is slightly larger than the size of the stage 42, and the rectangular hole is on the path of the main optical axis.

[0062] The mobile platform mounting plate 56 is a metal plate with a rectangular cross-section. The mobile platform mounting plate 56 spans the right support frame 21 and the left support frame 22. Both ends of the mobile platform mounting plate 56 are connected to the right support frame 21 and the left support frame 22 by screws. There are two mobile platform mounting plates 56, and the two mobile platform mounting plates 56 are distributed at intervals.

[0063] The lateral moving component 51 is the same as the longitudinal moving component 52, and both include a screw motor module 54 and a first linear slide pair 55, wherein the screw motor module 54 and the first linear slide pair 55 in the lateral moving component 51 are respectively fixed on two moving platform mounting plates 56, and extend laterally along the connecting plate 53, a moving block is installed on the screw of the screw motor module 54, and the moving block can move under the action of the screw, and the lower surface of the connecting plate 53 is fixed to the moving block in the screw motor module 54 in the lateral moving component 51 and the slider in the first linear slide pair 55 by screws, and under the action of the screw motor module 54 in the lateral moving component 51, the connecting plate 53 moves along the first linear slide pair 55.

[0064] The lead screw motor module 54 and the first linear slide rail pair 55 in the longitudinal moving component 52 are fixed to the upper surface of the connecting plate 53 and close to the longitudinal edge of the connecting plate 53 by means of screws. The two ends of the stage 42 are respectively connected to the moving block and the slider of the lead screw in the longitudinal moving component 52. Under the action of the longitudinal moving component 52, the stage 42 moves longitudinally to achieve the adjustment of the microscopic imaging field of the cell culture plate 41 placed in the stage 42.

[0065] like Figure 1 , 3As shown in FIGS. 11 and 12 , the cell culture component 4 includes a cell culture plate 41 for culturing cells and tissues, a loading platform 42 for placing the cell culture plate 41 , and a sealing frame 44 and a sealing cover 45 for providing a closed culture environment for the cell culture plate 41 , wherein the sealing frame 44 is mounted on the upper panel 11 , and the sealing cover 45 is detachably covered on the sealing frame 44 to seal the sealing frame 44 .

[0066] The stage 42 is a rectangular groove with a hollow bottom in the middle, in which a standard 96-well, 24-well or other specification cell culture plate 41 can be placed. In order to facilitate the control of the culture temperature of cells or tissues, a heating film and a temperature sensor are provided on the frame of the stage 42. The temperature of the cell culture plate 41 installed therein can be controlled by heat conduction to provide a suitable temperature for cell culture.

[0067] The cell culture plate 41 is an existing transparent porous cell culture plate, and the cell culture plate 41 has a plurality of cell culture chambers 43 for culturing cells or tissues. In this embodiment, a cell culture plate 41 with 24 cell culture chambers 43 is preferably used.

[0068] The sealing frame 44 is a rectangular frame with a certain wall thickness. The sealing frame 44 is fixed to the upper surface of the upper panel 11 and covers the outside of the functional hole 14. The cell culture plate 41 can be placed in the stage 42 through the sealing frame 44 and the functional hole 14.

[0069] The sealing cover 45 is a rectangular transparent plastic cover. The main function of the sealing cover 45 is to close the sealing frame 44 to provide a suitable gas environment for cell culture. In order to facilitate the sealing cover 45 to close the sealing frame 44, a long sealing frame groove 47 is concave on the inner side wall around the sealing frame 44, and at the same time, a long sealing cover protrusion 48 is correspondingly convex on the outer side wall around the sealing cover 45. The sealing cover protrusion 48 cooperates with the sealing frame groove 47 to achieve the sealing of the sealing frame 44 by the sealing cover 45. In addition, in order to ensure that it has a certain degree of sealing, a rubber sealing ring is also fixed on the contact surface between the sealing cover 45 and the sealing frame 44.

[0070] In order to provide a suitable gas environment for cell culture, two gas nozzle joints 46 are screwed on the outer wall of the sealing frame 44, which are connected to the external air pipe through the gas nozzle joints 46 to introduce suitable gas into the sealing frame 44 to provide a suitable gas environment for cell culture.

[0071] In order to ensure that the cell culture component 4 has a certain sealing performance, a flexible stage sealing ring 49 is placed on the upper surface of the stage 42. The stage sealing ring 49 fills the gap between the stage 42 and the upper panel 11 to ensure a certain sealing performance. In order to prevent the stage 42 from exceeding the functional hole 14 when the two-dimensional mobile platform 5 moves, the size of the stage 42 is larger than the size of the functional hole 14.

[0072] In this embodiment, regarding the control of the moving distance of each movable displacement structure, in addition to being controlled by the step number of the motor, a plurality of photoelectric switches can be set in the device to control the moving distance thereof.

[0073] like Figure 1 , 2 , 3 and 13, the bright field light source assembly 3 includes a bright field light source module 31, a lifting screw rod module 32, a support frame 33 and a support back plate 34. There are two support frames 33, which are fixed on the upper plate 11 in a left-right distribution. The support back plate 34 It is fixedly installed at the rear of the support frame 33 distributed on the left and right, the lifting screw module 32 is vertically fixedly installed on the support back plate 34, and the bright field light source module 31 is fixedly installed on the slider of the lifting screw module 32. By manually rotating the knob above the screw module, the slider can be driven to move up and down, thereby realizing the up and down movement of the bright field light source module 31 in the vertical direction and realizing the adjustment of the field of view brightness.

[0074] In addition, in order to protect the lifting screw module 32, a screw module housing 35 is wrapped around the lifting screw module 32, and the lower end of the screw module housing 35 is fixed to the upper panel 11, and the screw module housing 35 does not affect the normal use of the lifting screw module 32.

[0075] The bright field light source module 31 is an existing light source module and can be used directly.

[0076] A cell culture imaging method of a cell culture microscopy imaging integrated device of the present invention comprises the following steps: Cell and tissue culture: A1: Place cells, tissues and culture fluid to be cultured into the cell culture chamber 43 in the cell culture plate 41, and place them in multiple cell culture chambers 43 for comparison and reference; A2: Place the cell culture plate 41 on the stage 42 and cover it with a sealing cover 45 to seal the cell culture plate 41; A3: Open the air nozzle joint 46 to exhaust the air in the cell culture plate 41, and inject the specified gas into the cell culture plate 41, then turn on the heating film and set it to 37°C for heating, so as to provide a suitable gas environment and temperature environment for the cells in the cell culture component 4, and then carry out cell and tissue culture; Live bright-field microscopy: B1: Adjust the transmission mechanism in the fluorescence excitation assembly 8 to translate the fluorescence excitation module 801 as a whole to outside the main optical axis. At this time, the positions of the fluorescence excitation module 801 and the objective lens module 702 do not correspond. B2: Turn on the bright field light source module 31 and the camera 91; B3: Adjust the distance between the two shading plates 61 in the automatic shading assembly 6 to expose the cell culture plate 41. At this time, the imaging status can be viewed in real time through the display screen 15; B4: The intensity of the bright field light source module 31 is adjusted by the lifting screw module 32 to adjust the brightness of the field of view, and then the position of the cell culture plate 41 is adjusted by the two-dimensional mobile platform 5 to switch the observation field of view; B5: By adjusting the objective lens screw lifting module 701, the height of the objective lens module 702 in the vertical direction is adjusted to achieve focusing at different heights. The rotation of the objective lens turntable 705 is controlled by the turntable driving motor 711 to switch different objective lenses 713 to achieve observations with different magnifications. Real-time fluorescence microscopy: C1: Adjust the transmission mechanism in the fluorescence excitation assembly 8 to translate the fluorescence excitation module 801 as a whole to the main optical axis. At this time, the fluorescence excitation module 801 corresponds to the position of the objective lens module 702; C2: Turn on the bright field light source module 31 and the camera 91; C3: adjusting the distance between the two shading plates 61 in the automatic shading assembly 6 to expose the cell culture plate 41, and at this time, the imaging status can be viewed in real time through the display screen 15; C4: adjusting the intensity of the bright field light source module 31 through the lifting screw module 32 to adjust the brightness of the field of view, and then using the two-dimensional mobile platform 5 to adjust the position of the cell culture plate 41 to switch the observation field of view; C5: By adjusting the objective lens screw lifting module 701, the height of the objective lens module 702 in the vertical direction is adjusted to achieve focusing at different heights, and the rotation of the objective lens turntable 705 is controlled by the turntable driving motor 711 to switch different objective lenses 713 to achieve observations with different magnifications; C6: Adjust the transmission mechanism in the fluorescence excitation component 8 to move the fluorescence excitation module 801 containing different wavelengths to the main optical axis to achieve multi-channel fluorescence imaging.

[0077] The above embodiments are only for illustrating the technical idea of ​​the present invention, and cannot be used to limit the protection scope of the present invention. Any changes made on the basis of the technical solution in accordance with the technical idea proposed by the present invention shall fall within the protection scope of the present invention.

Claims

1. A cell culture microscopy imaging integrated device, characterized in that: include: A housing (1), wherein a display screen (15) is disposed on the housing (1), the display screen (15) controls the operation of the device and displays cell imaging in the device, and a support frame (2) for mounting other components in the device is disposed in the housing (1); A bright field light source assembly (3) is arranged at the top of the device, and the bright field light source assembly (3) provides illumination for cell microscopic imaging, and the brightness of the bright field light source assembly (3) is adjustable; A cell culture component (4) is arranged in the middle of the device and is located below the bright field light source component (3). Cells or tissues to be cultured are placed in a cell culture plate (41) in the cell culture component (4) for culture. The culture environment of the cells in the cell culture component (4) is adjustable. An objective lens assembly (7) is arranged on a bottom plate (13) in the housing (1) via a support frame (703) and is located below the cell culture assembly (4); the objective lens assembly (7) comprises an objective lens screw lifting module (701) and an objective lens module (702); the magnification and clarity of cell observation in the cell culture assembly (4) are adjusted via the objective lens module (702) and the objective lens screw lifting module (701); An automatic shading component (6) is arranged between the objective lens component (7) and the cell culture component (4), wherein the automatic shading component (6) comprises two shading plates (61), and the interval between the two shading plates (61) is adjustable; A fluorescence excitation component (8) is arranged below the objective lens component (7), the fluorescence excitation component (8) comprising a plurality of fluorescence excitation modules (801) and a fluorescence excitation module fixing plate (802), the fluorescence excitation modules (801) used for fluorescence microscopic imaging are arranged on the fluorescence excitation module fixing plate (802), and the upper and lower corresponding relationships between different fluorescence excitation modules (801) and the objective lens component (7) are adjustable; An imaging component (9) is arranged below the fluorescent excitation component (8) and is located at the bottom of the device. The imaging component (9) is connected to a display screen (15). The imaging component (9) photographs the cells observed in the cell culture component (4) through the objective lens component (7) and transmits the photographed image to the display screen (15) in real time.

2. The cell culture microscopy imaging integrated device according to claim 1, characterized in that: The bright field light source assembly (3) comprises a bright field light source module (31) and a lifting screw module (32); the bright field light source module (31) is arranged on the lifting screw module (32), and the interval between the bright field light source module (31) and the cell culture assembly (4) is adjusted by the lifting screw module (32).

3. The cell culture microscopy imaging integrated device according to claim 1, characterized in that: The cell culture component (4) comprises a cell culture plate (41), a stage (42), a sealing frame (44) and a sealing cover (45); the sealing frame (44) is arranged on the upper panel (11) of the housing (1); the stage (42) is connected to the two-dimensional movable platform (5); the lateral and longitudinal positions of the stage (42) are controllably adjustable by the two-dimensional movable platform (5); the cell culture plate (41) passes through the sealing frame (44) and is placed on the stage (42); and the sealing cover (45) is detachably sealed to seal the sealing frame (44).

4. The cell culture microscopy imaging integrated device according to claim 3, characterized in that: A heating film and a temperature sensor are arranged on the stage (42), the heating film providing a specified culture temperature for the cells in the cell culture component (4), and a gas nozzle joint (46) is arranged on the sealing frame (44), the gas nozzle joint (46) is connected to an external air pipe, and provides a specified gas environment for the cells in the cell culture component (4).

5. The cell culture microscopy imaging integrated device according to claim 1, characterized in that: The two-dimensional mobile platform (5) comprises a transverse moving component (51), a longitudinal moving component (52) and a mobile platform mounting plate (56); the transverse moving component (51) and the longitudinal moving component (52) are mounted on the support frame (2) via the mobile platform mounting plate (56); the object carrier (42) in the cell culture component (4) is arranged on the longitudinal moving component (52); and the position of the object carrier (42) is controllably adjustable via the two-dimensional mobile platform (5).

6. The cell culture microscopy imaging integrated device according to claim 1, characterized in that: The automatic shading assembly (6) comprises a front support plate (62) and a rear support plate (63), the front support plate (62) and the rear support plate (63) being arranged on the support frame (2), a slide groove (65) being arranged on the front support plate (62) and the rear support plate (63), two shading plates (61) being inserted into the slide groove (65) and arranged opposite to each other, a moving assembly being arranged on the shading plate (61), and the shading plate (61) being controllably slidable along the slide groove (65) by means of the moving assembly.

7. The cell culture microscopy imaging integrated device according to claim 1, characterized in that: The objective lens module (702) comprises an objective lens turntable (705) and an objective lens turntable base (706); a plurality of objective lenses (713) of different magnifications are arranged on the objective lens turntable (705); the objective lens turntable (705) is arranged on the objective lens turntable base (706); the objective lens turntable (705) is controllably rotated relative to the objective lens turntable base (706) by the drive of a turntable drive motor (711) and a gear set, thereby completing the switching of objective lenses (713) of different magnifications; A light-transmitting hole (710) is provided on the objective lens turntable base (706), and the correspondence between the plurality of objective lenses (713) and the light-transmitting holes (710) is adjusted by a turntable driving motor (711).

8. The cell culture microscopy imaging integrated device according to claim 1, characterized in that: There are multiple fluorescence excitation modules (801), and the wavelengths, excitation light filters (816) and emission light filters (819) of the multiple fluorescence excitation modules (801) are different.

9. The cell culture microscopy imaging integrated device according to claim 1, characterized in that: The imaging assembly (9) comprises a camera (91), a horizontal optical path pipe (92), a right-angle optical path block (95) and an imaging lens (96); one end of the horizontal optical path pipe (92) is connected to the camera (91), and the other end is connected to the right-angle optical path block (95); the imaging lens (96) is arranged above the right-angle optical path block (95); and light from above the imaging lens (96) is received by the CCD of the camera (91) via the imaging lens (96), the right-angle optical path block (95) and the horizontal optical path pipe (92).

10. The cell culture imaging method according to any one of claims 1 to 9, characterized in that The following steps are involved: Cell and tissue culture: A1: placing cells, tissues and culture fluid to be cultured into a cell culture chamber (43) in a cell culture plate (41), and placing cells, tissues and culture fluid into multiple cell culture chambers (43) for comparison and reference; A2: Place the cell culture plate (41) on the stage (42), cover it with a sealing cover (45), and seal the cell culture plate (41); A3: Open the air nozzle joint (46) to exhaust the air in the cell culture plate (41), and inject designated air into the cell culture plate (41), turn on the heating film and set it to 37°C for heating, so as to provide a suitable gas environment and temperature environment for the cells in the cell culture component (4), and then carry out cell and tissue culture; Live bright-field microscopy: B1: adjusting the transmission mechanism in the fluorescence excitation assembly (8) to translate the fluorescence excitation module (801) as a whole to outside the main optical axis, at which point the positions of the fluorescence excitation module (801) and the objective lens module (702) do not correspond; B2: Turn on the bright field light source module (31) and the camera (91); B3: adjusting the distance between the two shading plates (61) in the automatic shading assembly (6) to expose the cell culture plate (41), and at this time, the imaging status can be viewed in real time through the display screen (15); B4: adjusting the intensity of the bright field light source module (31) by means of the lifting screw module (32) to adjust the brightness of the field of view, and then using the two-dimensional moving platform (5) to adjust the position of the cell culture plate (41) to switch the observation field of view; B5: By adjusting the objective lens screw lifting module (701), the height of the objective lens module (702) in the vertical direction is adjusted to achieve focusing at different heights, and the rotation of the objective lens turntable (705) is controlled by the turntable driving motor (711) to switch different objective lenses (713) to achieve observation with different magnifications; Real-time fluorescence microscopy: C1: adjusting the transmission mechanism in the fluorescence excitation component (8) to translate the fluorescence excitation module (801) as a whole onto the main optical axis, at which point the fluorescence excitation module (801) and the objective lens module (702) are positioned correspondingly; C2: Turn on the bright field light source module (31) and the camera (91); C3: adjusting the distance between the two shading plates (61) in the automatic shading assembly (6) to expose the cell culture plate (41), and at this time, the imaging status can be viewed in real time through the display screen (15); C4: adjusting the intensity of the bright field light source module (31) by means of the lifting screw module (32) to adjust the brightness of the field of view, and then using the two-dimensional mobile platform (5) to adjust the position of the cell culture plate (41) to switch the observation field of view; C5: By adjusting the objective lens screw lifting module (701), the height of the objective lens module (702) in the vertical direction is adjusted to achieve focusing at different heights, and the rotation of the objective lens turntable (705) is controlled by the turntable driving motor (711) to switch different objective lenses (713) to achieve observation with different magnifications; C6: Adjust the transmission mechanism in the fluorescence excitation component (8) to move the fluorescence excitation module (801) containing different wavelengths to the main optical axis to achieve multi-channel fluorescence imaging.

Images