Multifunctional living cell imager

Abstract

The invention discloses a multifunctional living cell imager which is a full-automatic living cell imager with dual-purpose functions of laser scanning confocal imaging and wide-field fluorescence imaging and dual-purpose functions inside and outside a carbon dioxide incubator. The system comprises an in-box imaging unit, a multifunctional docking station and a computer. The in-box imaging unit comprises a bright field light source A, an objective table, an objective lens, a reflector A, an electric rotating wheel, an electric z-axis, a tube lens, a reflector B, an electric reflector, a camera, a confocal interface, a motion control unit A and a fluorescent light source. The multifunctional docking station comprises a motion control unit B, a bright field light source B, a laser confocal module, an electric objective table, a heat dissipation device and a carbon dioxide cover. The system gives consideration to in-box and out-box cell monitoring application, and is high in imaging speed, powerful in function and low in cost.

Description

Technical Field

[0001] The invention relates to the field of living cell imaging, and in particular to a multifunctional living cell imager, which is a fully automatic living cell imager with dual functions of laser scanning confocal imaging and wide-field fluorescence imaging, and dual functions of being used inside and outside a carbon dioxide incubator. Background Art

[0002] The carbon dioxide incubator provides the necessary conditions for constant temperature and humidity for living cell culture. In the process of cell culture, observation, photography and data processing at high resolution through an inverted desktop microscope are essential steps. There are some problems in this process: 1. When the cell culture container is moved from the incubator to the stage of the inverted desktop microscope outside for operation, the temperature and humidity of the cell culture environment and other parameters have all changed, which is not conducive to cell culture. The longer the operation time outside the box, the greater the impact on the cells, which is not conducive to control testing; 2. Although in recent years some companies have developed some in-box live cell imagers that are small enough to be placed in the incubator to monitor cells, they can already achieve bright field, phase contrast, wide field fluorescence and electric Z-axis autofocus and even XY electric scanning functions. However, these products that have been launched are subject to the requirement that the temperature in the incubator must be minimized, so the light source and camera with high heat generation inside the live cell imager cannot achieve continuous imaging and photography. 3. Existing in-box live cell imagers cannot achieve laser confocal scanning imaging, so they cannot achieve low-cost high-throughput and high-content screening functions.

[0003] Therefore, the present invention aims at the above three defects of the existing in-box live cell imager products on the market, and proposes a multifunctional live cell imager, which has the dual functions of laser scanning confocal imaging and wide-field fluorescence imaging, as well as the dual functions of inside and outside the carbon dioxide incubator. The present invention is expected to solve the above problems. The present invention includes an in-box imaging unit and a multifunctional docking station. The in-box imaging unit can be used to monitor cells in the incubator, and can also be used outside the incubator in combination with the multifunctional docking station. The light source and camera in the in-box imaging unit can work continuously by enhancing the heat dissipation capacity. The external carbon dioxide hood provided by the multifunctional docking station provides a stable temperature and humidity environment outside the box, which is conducive to long-term observation. In addition, the conventional transmission illumination column provided by the multifunctional docking station realizes the enhanced phase contrast imaging function. In addition, it can be connected to the laser scanning confocal module to realize the laser scanning confocal imaging function. Summary of the invention

[0004] The object of the present invention is to provide a multifunctional living cell imager, which is a fully automatic living cell imager having dual functions of laser scanning confocal imaging and wide-field fluorescence imaging and dual functions of being used inside and outside a carbon dioxide incubator.

[0005] To achieve the above-mentioned purpose, the present invention adopts the following technical scheme: a multifunctional living cell imager, comprising an in-box imaging unit, a multifunctional docking station and a computer; the in-box imaging unit comprises a bright field light source A, a stage, an objective lens, a reflector A, an electric rotating wheel, an electric z-axis, a tube lens, a reflector B, an electric reflector, a camera, a confocal interface, a motion control unit A, and a fluorescent light source; the multifunctional docking station comprises a motion control unit B, a bright field light source B, a laser confocal module, a motorized stage, a heat dissipation device and a carbon dioxide hood; In the built-in bright field and simple phase contrast imaging modes, the output end of the computer is connected to the input end of the motion control unit A, the other output end of the computer is connected to the input end of the camera, the output end of the motion control unit A is connected to the input end of the bright field light source A, the other output end of the motion control unit A is connected to the input end of the electric z-axis, another output end of the motion control unit A is connected to the input end of the electric rotating wheel, another output end of the motion control unit A is connected to the input end of the electric reflector, the output end of the bright field light source A is connected to the input end of the stage, the output end of the stage is connected to the input end of the objective lens, the output end of the electric z-axis is connected to the other input end of the objective lens, the output end of the objective lens is connected to the input end of the reflector A, the output end of the reflector A is connected to the other input end of the electric rotating wheel, the output end of the electric rotating wheel is connected to the input end of the tube mirror, the output end of the tube mirror is connected to the input end of the reflector B, the output end of the reflector B is connected to the other input end of the electric reflector, and the output end of the electric reflector is connected to the other input end of the camera; In the built-in epi-fluorescence imaging mode, the output end of the computer is connected to the input end of the motion control unit A, the other output end of the computer is connected to the input end of the camera, the output end of the motion control unit A is connected to the input end of the fluorescent light source, the other output end of the motion control unit A is connected to the input end of the electric z-axis, another output end of the motion control unit A is connected to the input end of the electric rotating wheel, another output end of the motion control unit A is connected to the input end of the electric reflecting mirror, the output end of the stage is connected to the input end of the objective lens, the output end of the electric z-axis is connected to the other input end of the objective lens, the output end of the objective lens is connected to the input end of the reflecting mirror A, the output end of the reflecting mirror A is connected to the other input end of the electric rotating wheel, the output end of the electric rotating wheel is connected to the input end of the tube mirror, the output end of the tube mirror is connected to the input end of the reflecting mirror B, the output end of the reflecting mirror B is connected to the other input end of the electric reflecting mirror, and the output end of the electric reflecting mirror is connected to the other input end of the camera; In the external enhanced phase contrast imaging mode, the output end of the computer is connected to the input end of the motion control unit A, the other output end of the computer is connected to the input end of the camera, another output end of the computer is connected to the input end of the motion control unit B, the output end of the motion control unit B is connected to the input end of the bright field light source B, the output end of the motion control unit A is connected to the input end of the electric z-axis, the other output end of the motion control unit A is connected to the input end of the electric rotating wheel, another output end of the motion control unit A is connected to the input end of the electric reflector, the output end of the bright field light source B is connected to the input end of the stage, the output end of the stage is connected to the input end of the objective lens, the output end of the electric z-axis is connected to the other input end of the objective lens, the output end of the objective lens is connected to the input end of the reflector A, the output end of the reflector A is connected to the other input end of the electric rotating wheel, the output end of the electric rotating wheel is connected to the input end of the tube mirror, the output end of the tube mirror is connected to the input end of the reflector B, the output end of the reflector B is connected to the other input end of the electric reflector, and the output end of the electric reflector is connected to the other input end of the camera; In the external epi-fluorescence imaging with carbon dioxide hood mode, the output end of the computer is connected to the input end of the motion control unit A, another output end of the computer is connected to the input end of the camera, another output end of the computer is connected to the input end of the motion control unit B, the output end of the motion control unit A is connected to the input end of the fluorescent light source, another output end of the motion control unit A is connected to the input end of the electric z-axis, another output end of the motion control unit A is connected to the input end of the electric turntable, another output end of the motion control unit A is connected to the input end of the electric reflector, the output end of the stage is connected to the input end of the objective lens, the output end of the electric z-axis is connected to another input end of the objective lens, and the output end of the objective lens connected to the input end of the reflector A, the output end of the reflector A is connected to the other input end of the electric wheel, the output end of the electric wheel is connected to the input end of the tube mirror, the output end of the tube mirror is connected to the input end of the reflector B, the output end of the reflector B is connected to the other input end of the electric reflector, the output end of the electric reflector is connected to the other input end of the camera, the output end of the motion control unit B is connected to the input end of the carbon dioxide mask, the output end of the motion control unit B is connected to the input end of the heat sink, the output end of the heat sink is connected to the other input end of the fluorescent light source, the other output end of the heat sink is connected to another input end of the camera, and the output end of the fluorescent light source is connected to the other input end of the electric wheel; In the external laser confocal imaging and XY electric stage mode, the output end of the computer is connected to the input end of the motion control unit A, the other output end of the computer is connected to the input end of the laser confocal module, another output end of the computer is connected to the input end of the motion control unit B, the output end of the motion control unit B is connected to the input end of the electric stage, the output end of the motion control unit A is connected to the input end of the electric z-axis, the other output end of the motion control unit A is connected to the input end of the electric rotary wheel, another output end of the motion control unit A is connected to the input end of the electric reflector, the output end of the stage is connected to the input end of the objective lens, the output end of the electric z-axis is connected to the other input end of the objective lens, the output end of the objective lens is connected to the input end of the reflector A, the output end of the reflector A is connected to the other input end of the electric rotary wheel, the output end of the electric rotary wheel is connected to the input end of the tube mirror, the output end of the tube mirror is connected to the input end of the reflector B, the output end of the reflector B is connected to the other input end of the electric reflector, the output end of the electric reflector is connected to the input end of the confocal interface, and the output end of the confocal interface is connected to the other input end of the laser confocal module.

[0006] The working principle of the present invention is that compared with the existing in-box live cell imagers, the present invention provides multiple imaging function modes. In the built-in bright field and simple phase contrast imaging modes, the bright field light source A of the present invention is a foldable and retractable pop-up annular light source, which is located at a specific height directly above the stage. The divergent annular illumination light passes through the cell sample and interacts with the built-in phase contrast ring of the objective lens to produce a certain three-dimensional shadow effect. The light collected by the objective lens is reflected by the reflector A, passes through the empty space of the electric wheel, is focused by the tube lens, and then reflected by the reflector B and finally focused on the photoelectric photosensor unit of the camera to form an image. The computer receives the signal from the camera and controls the brightness of the bright field light source A through the motion control unit A. The electric z-axis equipped with the objective lens performs automatic focusing, and the electric wheel rotates and the electric reflector switches between two positions. In the epi-fluorescence imaging mode, the fluorescent light source of the present invention comprises a plurality of excitation light sources of different wavelengths, the generated excitation light is first collimated by a collimating lens, and then focused on the rear focal plane of the objective lens by a field lens for realizing Kohler illumination, the excitation light passes through an excitation filter in an electric rotating wheel, is reflected by a dichroic mirror, and then is reflected by a reflector A, the excitation light is irradiated on the sample to generate fluorescence, the fluorescence collected by the objective lens is reflected by the reflector A, passes through the dichroic mirror and the emission filter of the electric rotating wheel, is focused by a tube lens, and then is reflected by a reflector B and finally focused on a photoelectric photosensitive unit of a camera to form an image, a computer receives a signal from the camera and controls the brightness of the bright field light source A through a motion control unit A, and an electric z-axis equipped with an objective lens is operated Automatic focusing, the rotation of the electric wheel and the switching of the electric reflector to two positions; in the external enhanced phase contrast imaging mode, the bright field light source B of the present invention is a conventional transmission illumination column for an inverted desktop microscope, and the bright field light source B comprises a light emitter such as various gas discharge lamps or LED lamps, a condenser, a phase contrast plate and a field lens, and generates a divergent annular illumination light which passes through the cell sample and then interacts with the built-in phase contrast ring of the objective lens to produce a certain three-dimensional shadow effect, and the light collected by the objective lens is reflected by the reflector A, passes through the empty position of the electric wheel, is focused by the tube lens, and then reflected by the reflector B and finally focused on the photoelectric photosensitive unit of the camera to form an image, and the computer receives the signal of the camera and controls the bright field light source B through the motion control unit B The brightness of the image is adjusted, the electric z-axis with the objective lens is automatically focused, the electric wheel rotates and the electric reflector switches between two positions; in the external epi-fluorescence imaging with carbon dioxide hood mode, the difference compared with the built-in epi-fluorescence imaging mode is that the computer controls the heat dissipation device through the motion control unit B to force the heat dissipation of the fluorescent light source and the camera, so that the fluorescent light source and the camera can work in the continuous shooting mode. The motion control unit B also controls the gas mixing device, heating device, heat dissipation device and humidification device in the carbon dioxide hood to achieve the stability of the culture environment in the carbon dioxide hood, and can realize long-term cell culture and monitoring outside the box. The similarities compared with the built-in epi-fluorescence imaging mode are not repeated here;In the external laser confocal imaging and XY electric stage mode, the computer controls the electric z-axis through the motion control unit A to achieve automatic focusing, controls the electric wheel to be in the empty position, controls the electric reflector to switch to the confocal interface direction instead of the camera direction, and controls the electric stage in the XY direction through the motion control unit A to move the sample to achieve imaging at different positions. The computer controls the LED light source in the laser confocal module to generate excitation light, which passes through the dichroic mirror and the scanning galvanometer, and then passes through the electric reflector, reflector B, tube mirror, the empty position of the electric wheel, and reflector A, and then is focused on the sample by the objective lens to excite fluorescence. The fluorescence is then collected by the objective lens and passes through the reflector A, the empty position of the electric wheel, tube mirror, reflector B, electric reflector, and then reflected again by the scanning galvanometer, and then enters the photodetector through the dichroic mirror and the pinhole to generate an electrical signal. As the scanning galvanometer moves, the image is scanned point by point. The electrical signal can be processed by the computer to form a two-dimensional or three-dimensional image. ;

[0007] Furthermore, the bright field light source A includes a foldable support frame and a ring-shaped LED lamp. The foldable support frame can be flipped sideways and stored in the box of the multifunctional living cell imager. The ring-shaped LED lamp is used to generate ring-shaped illumination light, which is collected by the objective lens and then focused on the phase contrast ring in the objective lens to produce a three-dimensional shadow effect.

[0008] Furthermore, the bright field light source B comprises a gas discharge lamp or LED light source, a collimating lens, a phase difference ring plate and a condenser lens which are sequentially arranged along the direction of the light path, and the spacing between the condenser lens and the phase difference ring plate is adjustable.

[0009] Furthermore, the stage is a fixed stage, which is located on a part of the upper surface of the housing of the multifunctional living cell imager.

[0010] Furthermore, the objective lens is a phase contrast objective lens.

[0011] Furthermore, the electric rotating wheel is a circular, rectangular or polygonal rotating wheel, which is driven by a motor to rotate. Multiple fluorescent filter blocks are arranged in the electric rotating wheel for fluorescent imaging and an empty position for bright field imaging and confocal imaging. Each fluorescent filter block has an excitation filter, a dichroic mirror and an emission filter.

[0012] Furthermore, the electric z-axis is used to drive the objective lens module to adjust its position in the vertical direction.

[0013] Furthermore, the electric reflector is driven to rotate by a motor, the rotation axis of the motor is perpendicular to the optical path, and switches between two positions, one position is 45 degrees for reflecting light 90 degrees into the camera, and the other position is 90 degrees for passing the light beam to the confocal interface.

[0014] Furthermore, the motion control unit A and the motion control unit B have motor control function, analog input, analog output, digital input and digital output functions, and are connected to the computer via a network cable or RS485 or USB interface.

[0015] Furthermore, the fluorescent light source comprises a plurality of fluorescent channels, each of which comprises a gas discharge lamp or an LED light source, a collimating lens and a dichroic mirror arranged in sequence along the direction of the light path.

[0016] Furthermore, the heat dissipation device includes a heat sink and a fan. The heat sink is in close contact with the fluorescent light source and the camera. The airflow generated by the fan blows through the heat sink to take away the heat so that the temperature of the fluorescent light source and the camera will not be too high.

[0017] Furthermore, the confocal interface is a standard C-mount interface, which is used to connect a multifunctional living cell imager and a laser scanning confocal module.

[0018] Furthermore, the laser scanning confocal module includes an LED light source, a scanning galvanometer, a dichroic mirror, a pinhole and a photodetector. The LED light source includes an LED lamp, a collimating lens and an excitation filter. The scanning galvanometer is a mechanical device based on the electromagnetic actuation principle or the comb transducer electric actuation principle. The surface has a high reflectivity coating and is used to deflect the light beam in the XY bidirectional direction at high speed.

[0019] Furthermore, the carbon dioxide hood comprises a gas mixing device, a heating device, a heat dissipation device and a humidifying device to achieve a stable culture environment in the carbon dioxide hood, and can realize long-term cell culture and monitoring outside the box. BRIEF DESCRIPTION OF THE DRAWINGS

[0020] FIG1 is a schematic diagram showing the principle of working mode 1 of the present invention; FIG2 is a schematic diagram showing the principle of working mode 2 of the present invention; FIG3 is a schematic diagram of the working principle of the third working mode of the present invention; FIG4 is a schematic diagram showing the principle of working mode 4 of the present invention; FIG5 is a schematic diagram showing the principle of working mode 5 of the present invention; FIG6 is a schematic structural diagram of the electric wheel of the present invention; Figure 7 It is a schematic diagram of the internal structure of the imaging unit in the box of the present invention; Figure 8 It is a schematic diagram of the external structure of the imaging unit in the box of the present invention; Fig. 9 It is a schematic diagram of the connection between the imaging unit in the box and the multi-functional expansion dock of the present invention. DETAILED DESCRIPTION

[0021] The following is a further detailed description through specific implementation methods: The reference numerals in the drawings of the specification include: bright field light source A1, stage 2, objective lens 3, reflector A4, electric wheel 5, electric z-axis 6, tube lens 7, reflector B8, electric reflector 9, camera 10, confocal interface 11, motion control unit A12, computer 13, motion control unit B14, bright field light source B15, fluorescent light source 16, laser confocal module 17, electric stage 18, heat dissipation device 19 and carbon dioxide hood 20. The working method is the same as that of Figure 1 As shown: the output end of the computer 13 is connected to the input end of the motion control unit A12, the other output end of the computer 13 is connected to the input end of the camera 10, the output end of the motion control unit A12 is connected to the input end of the bright field light source A1, the other output end of the motion control unit A12 is connected to the input end of the electric z-axis 6, another output end of the motion control unit A12 is connected to the input end of the electric rotating wheel 5, another output end of the motion control unit A12 is connected to the input end of the electric reflecting mirror 9, the output end of the bright field light source A1 is connected to the input end of the stage 2, the output end of the stage 2 is connected to the input end of the objective lens 3, the output end of the electric z-axis 6 is connected to the other input end of the objective lens 3, the output end of the objective lens 3 is connected to the input end of the reflecting mirror A4, the output end of the reflecting mirror A4 is connected to the other input end of the electric rotating wheel 5, the output end of the electric rotating wheel 5 is connected to the input end of the tube mirror 7, and the output end of the tube mirror 7 is connected to the input end of the tube mirror 7. The output end is connected to the input end of the reflector B8, the output end of the reflector B8 is connected to another input end of the electric reflector 9, and the output end of the electric reflector 9 is connected to another input end of the camera 10. The bright field light source A1 of the present invention is a foldable and retractable pop-up annular light source, which is located at a specific height directly above the stage 2. The divergent annular illumination light passes through the cell sample and interacts with the built-in phase contrast ring of the objective lens 3 to produce a certain three-dimensional shadow effect. The light collected by the objective lens 3 is reflected by the reflector A4, passes through the empty space of the electric rotating wheel 5, is focused by the tube lens 7, and then reflected by the reflector B8 and finally focused on the photoelectric photosensitive unit of the camera 10 to form an image. The computer 13 receives the signal of the camera 10 and controls the brightness of the bright field light source A1 through the motion control unit A12. The electric z-axis 6 equipped with the objective lens 3 performs automatic focusing, and the rotation of the electric rotating wheel 5 and the electric reflector 9 switch between two positions.Working mode 2 is shown in Figure 2: the output end of the computer 13 is connected to the input end of the motion control unit A12, the other output end of the computer 13 is connected to the input end of the camera 10, the output end of the motion control unit A12 is connected to the input end of the fluorescent light source 16, the other output end of the motion control unit A12 is connected to the input end of the electric z-axis 6, another output end of the motion control unit A12 is connected to the input end of the electric rotating wheel 5, another output end of the motion control unit A12 is connected to the input end of the electric reflecting mirror 9, the output end of the stage 2 is connected to the input end of the objective lens 3, the output end of the electric z-axis 6 is connected to the other input end of the objective lens 3, the output end of the objective lens 3 is connected to the input end of the reflecting mirror A4, the output end of the reflecting mirror A4 is connected to the other input end of the electric rotating wheel 5, the output end of the electric rotating wheel 5 is connected to the input end of the tube mirror 7, the output end of the tube mirror 7 is connected to the input end of the reflecting mirror B8, and the output end of the reflecting mirror B8 is connected to the input end of the tube mirror 7. The end is connected to the other input end of the electric reflector 9, and the output end of the electric reflector 9 is connected to the other input end of the camera 10. The fluorescent light source 16 of the present invention includes multiple groups of excitation light sources with different wavelengths. The generated excitation light is first collimated by a collimating lens, and then focused on the rear focal plane of the objective lens 3 by the field lens that realizes Kohler illumination. The excitation light passes through the excitation filter in the electric rotating wheel 5, is reflected by the dichroic mirror, and then reflected by the reflector A. The excitation light is irradiated on the sample to generate fluorescence. The fluorescence collected by the objective lens 3 is reflected by the reflector A4, passes through the dichroic mirror and the emission filter of the electric rotating wheel 5, is focused by the tube lens 7, and then reflected by the reflector B8 and finally focused on the photoelectric photosensitive unit of the camera 10 to form an image. The computer 13 receives the signal of the camera 10 and controls the brightness of the bright field light source A1 through the motion control unit A12. The electric z-axis 6 equipped with the objective lens 3 performs automatic focusing, and the rotation of the electric rotating wheel 5 and the electric reflector 9 switch between two positions.Working mode three is shown in Figure 3: the output end of the computer 13 is connected to the input end of the motion control unit A12, another output end of the computer 13 is connected to the input end of the camera 10, another output end of the computer 13 is connected to the input end of the motion control unit B14, the output end of the motion control unit B14 is connected to the input end of the bright field light source B15, the output end of the motion control unit A12 is connected to the input end of the electric z-axis 6, another output end of the motion control unit A12 is connected to the input end of the electric rotating wheel 5, another output end of the motion control unit A12 is connected to the input end of the electric reflecting mirror 9, the output end of the bright field light source B15 is connected to the input end of the stage 2, the output end of the stage 2 is connected to the input end of the objective lens 3, the output end of the electric z-axis 6 is connected to the other input end of the objective lens 3, the output end of the objective lens 3 is connected to the input end of the reflecting mirror A4, the output end of the reflecting mirror A4 is connected to the other input end of the electric rotating wheel 5, and the output end of the electric rotating wheel 5 is connected to the input end of the tube mirror 7. The output end of the tube lens 7 is connected to the input end of the reflector B8, the output end of the reflector B8 is connected to the other input end of the electric reflector 9, the output end of the electric reflector 9 is connected to the other input end of the camera 10, the bright field light source B15 of the present invention is a conventional transmission lighting column for an inverted desktop microscope, the bright field light source B15 comprises a light emitter such as various gas discharge lamps or LED lamps, a condenser, a phase contrast plate and a field lens, and generates a divergent annular illumination light which passes through the cell sample and then interacts with the built-in phase contrast ring of the objective lens 3 to produce a certain three-dimensional shadow effect, the light collected by the objective lens 3 is reflected by the reflector A4, passes through the empty space of the electric rotating wheel 5, is focused by the tube lens 7, and then is reflected by the reflector B8 and finally focused on the photoelectric photosensitive unit of the camera 10 to form an image, the computer 13 receives the signal of the camera 10 and controls the brightness of the bright field light source B15 through the motion control unit B14, the electric z-axis 6 equipped with the objective lens 3 performs automatic focusing, the rotation of the electric rotating wheel 5 and the electric reflector 9 switch between two positions;Working mode 4 is shown in Figure 4: the output end of the computer 13 is connected to the input end of the motion control unit A12, another output end of the computer 13 is connected to the input end of the camera 10, another output end of the computer 10 is connected to the input end of the motion control unit B14, the output end of the motion control unit A12 is connected to the input end of the fluorescent light source 16, another output end of the motion control unit A12 is connected to the input end of the electric z-axis 6, another output end of the motion control unit A12 is connected to the input end of the electric rotating wheel 5, another output end of the motion control unit A12 is connected to the input end of the electric reflector 9, the output end of the stage 2 is connected to the input end of the objective lens 3, the output end of the electric z-axis 6 is connected to another input end of the objective lens 3, the output end of the objective lens 3 is connected to the input end of the reflector A4, the output end of the reflector A4 is connected to another input end of the electric rotating wheel 5, the output end of the electric rotating wheel 5 is connected to the input end of the tube mirror 7, the output end of the tube mirror 7 is connected to the input end of the reflector B8, and the output end of the reflector B8 is connected to another input end of the electric reflector 9. An input end is connected, an output end of the electric reflector 9 is connected to another input end of the camera 10, an output end of the motion control unit B14 is connected to an input end of the carbon dioxide hood 20, an output end of the motion control unit B14 is connected to an input end of the heat sink 19, an output end of the heat sink 19 is connected to another input end of the fluorescent light source 16, another output end of the heat sink 19 is connected to another input end of the camera 10, and an output end of the fluorescent light source 16 is connected to another input end of the electric turntable 5. The difference compared with the working mode 2 is that the computer 13 controls the heat sink 19 through the motion control unit B14 to force the fluorescent light source 16 and the camera 10 to dissipate heat, so that the fluorescent light source 16 and the camera 10 can work in a continuous shooting mode. The motion control unit B14 also controls the gas mixing device, heating device, heat sink and humidification device in the carbon dioxide hood 20 to achieve the stability of the culture environment in the carbon dioxide hood 20, and can realize long-term cell culture and monitoring outside the box. The same as the working mode 2 is that it is not repeated here;Working mode 5 is shown in FIG5 : the output end of the computer 13 is connected to the input end of the motion control unit A12, another output end of the computer 13 is connected to the input end of the laser confocal module 17, another output end of the computer 13 is connected to the input end of the motion control unit B14, the output end of the motion control unit B14 is connected to the input end of the electric stage 18, the output end of the motion control unit A12 is connected to the input end of the electric z-axis 6, another output end of the motion control unit A12 is connected to the input end of the electric wheel 5, and the motion control unit A12 Another output end is connected to the input end of the electric reflector 9, the output end of the stage 2 is connected to the input end of the objective lens 3, the output end of the electric z-axis 6 is connected to the other input end of the objective lens 3, the output end of the objective lens 3 is connected to the input end of the reflector A4, the output end of the reflector A4 is connected to the other input end of the electric wheel 5, the output end of the electric wheel 5 is connected to the input end of the tube lens 7, the output end of the tube lens 7 is connected to the input end of the reflector B8, the output end of the reflector B8 is connected to the other input end of the electric reflector 9, and the output end of the electric reflector 9 is connected to the confocal interface The input end of the confocal interface 11 is connected to the input end of the laser confocal module 17, the output end of the confocal interface 11 is connected to the other input end of the laser confocal module 17, the computer 13 controls the electric z-axis 6 to achieve automatic focusing through the motion control unit A12, controls the electric wheel 5 to be located in the empty position, controls the electric reflector 9 to switch to the direction of the confocal interface 11 instead of the camera 10, the computer 3 controls the electric stage 18 in the XY direction through the motion control unit A12 to move the sample to achieve imaging at different positions, and the computer 3 controls the LED light source in the laser confocal module 17 to generate excitation light , after being reflected by the dichroic mirror and the scanning galvanometer, it passes through the electric reflector 9, the reflector B9, the tube mirror 7, the empty space of the electric wheel 5, and the reflector A4, and then is focused on the sample by the objective lens 3 to excite fluorescence. The fluorescence is then collected by the objective lens 3 and passes through the reflector A4, the empty space of the electric wheel 5, the tube mirror 7, the reflector B8, and the electric reflector 9. After being reflected again by the scanning galvanometer, it passes through the dichroic mirror and the pinhole and enters the photodetector to generate an electrical signal. As the scanning galvanometer moves, it scans point by point to form an image. The electrical signal can be processed by the computer 13 to form a two-dimensional or three-dimensional image. ;

[0022] Furthermore, the bright field light source A1 includes a foldable support frame and a ring-shaped LED lamp. The foldable support frame can be flipped sideways and stored in the box of the multifunctional living cell imager. The ring-shaped LED lamp is used to generate a ring-shaped illumination light which is collected by the objective lens 3 and then focused on the phase contrast ring in the objective lens 3 to produce a three-dimensional shadow effect.

[0023] Furthermore, the bright field light source B15 includes a gas discharge lamp or LED light source, a collimating lens, a phase difference ring plate and a focusing lens which are sequentially arranged along the direction of the light path, and the distance between the focusing lens and the phase difference ring plate is adjustable.

[0024] Furthermore, the stage 2 is a fixed stage, which is located on a part of the upper surface of the housing of the multifunctional living cell imager.

[0025] Furthermore, the objective lens 3 is a phase contrast objective lens.

[0026] Furthermore, the electric rotating wheel 5 is a circular, rectangular or polygonal rotating wheel, which is driven by a motor to rotate. A plurality of fluorescent filter blocks are arranged in the electric rotating wheel for fluorescent imaging and an empty position for bright field imaging and confocal imaging. Each fluorescent filter block has an excitation filter, a dichroic mirror and an emission filter.

[0027] Furthermore, the electric z-axis 6 is used to drive the objective lens 3 module to adjust its position in the vertical direction.

[0028] Furthermore, the electric reflector 9 is driven to rotate by a motor, the rotation axis of the motor is perpendicular to the optical path, and switches between two positions, one position is 45 degrees for reflecting light 90 degrees into the camera 10, and the other position is 90 degrees for passing the light beam to the confocal interface 11.

[0029] Furthermore, the motion control unit A12 and the motion control unit B14 have motor control function, analog input, analog output, digital input and digital output functions, and are connected to the computer 13 via a network cable or RS485 or USB interface.

[0030] Furthermore, the fluorescent light source 16 comprises a plurality of fluorescent channels, each of which comprises a gas discharge lamp or an LED light source, a collimating lens and a dichroic mirror arranged in sequence along the direction of the light path.

[0031] Furthermore, the heat dissipation device 19 includes a heat sink and a fan. The heat sink is in close contact with the fluorescent light source 16 and the camera 10. The airflow generated by the fan blows through the heat sink to take away the heat so that the temperature of the fluorescent light source 16 and the camera 10 will not be too high.

[0032] Furthermore, the confocal interface 11 is a standard C-mount interface, which is used to connect the multifunctional living cell imager and the laser scanning confocal module 17 .

[0033] Furthermore, the laser scanning confocal module 17 includes an LED light source, a scanning galvanometer, a dichroic mirror, a pinhole and a photodetector. The LED light source includes an LED lamp, a collimating lens and an excitation filter. The scanning galvanometer is a mechanical device based on the electromagnetic actuation principle or the comb transducer electric actuation principle, and has a high reflectivity coating on the surface, which is used to deflect the light beam in the XY bidirectional direction at high speed.

[0034] Furthermore, the carbon dioxide hood 20 comprises a gas mixing device, a heating device, a heat dissipation device and a humidifying device to achieve a stable culture environment in the carbon dioxide hood 20, and can realize long-term cell culture and monitoring outside the box.

[0035] The above is only an embodiment of the present invention. The common sense such as the known specific structure and characteristics in the scheme is not described in detail here. The ordinary technicians in the relevant field know all the common technical knowledge in the technical field of the invention before the application date or priority date, can obtain all the existing technologies in the field, and have the ability to apply the conventional experimental means before that date. The ordinary technicians in the relevant field can improve and implement this scheme in combination with their own abilities under the enlightenment given by this application. Some typical known structures or known methods should not become obstacles for ordinary technicians in the relevant field to implement this application. It should be pointed out that for those skilled in the art, without departing from the structure of the present invention, several deformations and improvements can be made, especially the embodiments of the present invention can be enlarged or reduced in proportion, which should also be regarded as the protection scope of the present invention, which will not affect the effect of the implementation of the present invention and the practicality of the patent. The protection scope required by this application shall be based on the content of its claims, and the specific implementation methods and other records in the specification can be used to interpret the content of the claims.

Claims

1. A multifunctional living cell imaging instrument, characterized in that: It includes an in-box imaging unit, a multifunctional expansion dock and a computer; the in-box imaging unit includes a bright field light source A, a stage, an objective lens, a reflector A, an electric rotating wheel, an electric z-axis, a tube mirror, a reflector B, an electric reflector, a camera, a confocal interface, a motion control unit A, and a fluorescent light source; the multifunctional expansion dock includes a motion control unit B, a bright field light source B, a laser confocal module, a motorized stage, a heat dissipation device and a carbon dioxide hood.

2. A multifunctional living cell imaging device as claimed in claim 1, characterized in that: The bright field light source A comprises a foldable support frame and an annular LED lamp. The foldable support frame can be flipped sideways and stored in the box of the multifunctional living cell imager. The annular LED lamp is used to generate annular illumination light which is collected by the objective lens and then focused on the phase contrast ring in the objective lens to produce a three-dimensional shadow effect.

3. A multifunctional living cell imaging device as claimed in claim 1, characterized in that: The bright field light source B comprises a gas discharge lamp or LED light source, a collimating lens, a phase difference ring plate and a condenser lens which are sequentially arranged along the direction of the light path, and the distance between the condenser lens and the phase difference ring plate is adjustable.

4. The multifunctional living cell imaging device according to claim 1, characterized in that: The electric rotating wheel is a circular, rectangular or polygonal rotating wheel, which is driven by a motor to rotate. Multiple fluorescent filter blocks are arranged in the electric rotating wheel for fluorescent imaging and an empty position for bright field imaging and confocal imaging. Each fluorescent filter block has an excitation filter, a dichroic mirror and an emission filter.

5. The multifunctional living cell imaging device according to claim 1, characterized in that: The electric z-axis is used to drive the objective lens module to adjust its position in the vertical direction.

6. The multifunctional living cell imaging device according to claim 1, characterized in that: The electric reflector is driven to rotate by a motor, the rotation axis of the motor is perpendicular to the optical path, and switches between two positions, one position is 45 degrees for reflecting light 90 degrees into the camera, and the other position is 90 degrees for passing the light beam to the confocal interface.

7. The multifunctional living cell imaging device according to claim 1, characterized in that: The motion control unit A and the motion control unit B have motor control function, analog input, analog output, digital input and digital output functions, and are connected to the computer via a network cable or RS485 or USB interface.

8. The multifunctional living cell imaging device according to claim 1, characterized in that: The fluorescent light source comprises a plurality of fluorescent channels, each of which has a gas discharge lamp or LED light source, a collimating lens and a dichroic mirror arranged in sequence along the direction of the light path.

9. The multifunctional living cell imaging device according to claim 1, characterized in that: The heat dissipation device comprises a heat sink and a fan. The heat sink is in close contact with the fluorescent light source and the camera. The airflow generated by the fan blows through the heat sink to take away the heat so that the temperature of the fluorescent light source and the camera will not be too high.

10. The multifunctional living cell imaging device according to claim 1, characterized in that: The laser scanning confocal module includes an LED light source, a scanning galvanometer, a dichroic mirror, a pinhole and a photodetector. The LED light source includes an LED lamp, a collimating lens and an excitation filter. The scanning galvanometer is a mechanical device based on the electromagnetic actuation principle or the comb transducer electric actuation principle. The surface has a high reflectivity coating and is used for high-speed XY bidirectional deflection of the light beam.

11. The multifunctional living cell imaging device according to claim 1, characterized in that: The carbon dioxide hood comprises a gas mixing device, a heating device, a heat dissipation device and a humidifying device to achieve a stable culture environment in the carbon dioxide hood, and can realize long-term cell culture and monitoring outside the box.

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