Microscopic imaging analysis device for living cells under continuous or discontinuous sample adding condition

Abstract

The invention discloses a microimaging analysis device for living cells under a continuous or discontinuous sample adding condition, which belongs to the technical field of cell analysis devices, and comprises an incubator, a clamping mechanism, a culture dish, a top cover, a continuous sample adding mechanism and a microscope, the continuous sample adding mechanism comprises two mounting frames, two pulling assemblies, two fixing plates, two rotating rings, two rotating assemblies, a liquid inlet pipe and a liquid outlet pipe, when the top cover is taken down, when continuous sample adding of the medicine is needed, the pulling assemblies are started, the fixing plates are driven to translate, after the ends of the liquid inlet pipe and the liquid outlet pipe are located above a culture dish, the rotating assemblies are started, and the liquid inlet pipe and the liquid outlet pipe are driven to translate; the liquid inlet pipe and the liquid outlet pipe are driven by the rotating ring to rotate, so that the end parts of the liquid inlet pipe and the liquid outlet pipe obliquely rotate into the culture dish, and the liquid inlet pipe and the liquid outlet pipe are matched to complete continuous sample adding of the medicine, so that continuous sample adding of the medicine can be realized by adopting the structure; the method is suitable for drug mechanism research and application of a long-time living cell imaging technology, and is higher in universality.

Description

Technical Field

[0001] The invention belongs to the technical field of cell analysis devices and provides a microscopic imaging analysis device for living cells under continuous or intermittent sample addition conditions. Background Art

[0002] Real-time recording of cell culture has become one of the important experimental techniques for studying cells and related disciplines, especially stem cell research. It has the advantages of being intuitive, easy to control, and convenient for exploring the phenomena and mechanisms of cell growth, division, differentiation, transformation, stress response, signal transduction, regulation and control, aging, apoptosis, damage, necrosis, etc. Currently, most of the real-time recording of cell culture on the market adopts a combination of a microenvironment control system and an inverted microscope system.

[0003] When performing microscopic imaging analysis on living cells during culture, drugs need to be added to the living cell culture dish, but existing living cell microscopic imaging analysis devices cannot achieve continuous drug addition and have poor versatility. Summary of the invention

[0004] In view of this, the purpose of the present invention is to provide a microscopic imaging analysis device for living cells under continuous or intermittent sample addition conditions, aiming to solve the problem that existing living cell microscopic imaging analysis devices cannot achieve continuous drug addition and have poor versatility.

[0005] To achieve the above object, the present invention provides the following technical solutions:

[0006] The present invention provides a microscopic imaging analysis device for living cells under continuous or intermittent sample addition conditions, the microscopic imaging analysis device for living cells under continuous or intermittent sample addition conditions comprises a culture box, a clamping mechanism, a culture dish, a top cover, a continuous sample addition mechanism and a microscope, the culture dish is fixed inside the culture box through the clamping mechanism, the top of the culture box is provided with a detachable top cover, and the microscope is installed on the outside of the culture box;

[0007] The continuous loading mechanism includes two mounting frames, two pulling components, two fixed plates, two rotating rings, two rotating components, a liquid inlet pipe and a liquid outlet pipe. The two mounting frames are respectively mounted on both sides of the outside of the incubator, and the fixed plate is mounted on one side of each mounting frame facing the incubator through the pulling component. The rotating ring is rotatably arranged at the center of each fixed plate, and the liquid inlet pipe and the liquid outlet pipe are respectively embedded in the interiors of the two rotating rings. The rotating component is arranged on each fixed plate, and the output end of the rotating component is mechanically transmitted with the rotating ring. The liquid inlet pipe and the liquid outlet pipe both pass through the outer wall of the incubator, and the liquid inlet pipe and the liquid outlet pipe are both arranged in an L-shaped structure. The ends of the liquid inlet pipe and the liquid outlet pipe located inside the incubator correspond to the culture dish.

[0008] Each of the pulling components is composed of two electric telescopic rods, and the electric telescopic rods are arranged on both sides of each of the fixing plates, and one end of each of the electric telescopic rods away from the fixing plate is connected to the mounting frame.

[0009] Wherein, each of the rotating components includes a driving motor, an output gear and a transmission gear. The driving motor is installed on the fixed plate. The output end of the driving motor is provided with the output gear. The transmission gear is installed on the outside of the rotating ring, and the output gear is meshed with the transmission gear.

[0010] Wherein, two limiting rings are arranged outside each rotating ring, and the two limiting rings are respectively located on two sides of the corresponding fixing plate.

[0011] The clamping mechanism comprises two clamping cylinders and two arc-shaped clamping plates. The clamping cylinders are arranged on both sides of the interior of the incubator. The output end of each clamping cylinder is provided with the arc-shaped clamping plate, and the arc-shaped clamping plate corresponds to the culture dish.

[0012] The incubator is also provided with a water tank and a temperature regulating module inside, and the temperature regulating module is used to adjust the ambient temperature inside the incubator. The outer wall of the incubator is also provided with a water inlet and a CO 2 The water inlet corresponds to the water tank, and the CO 2 An intermittent fine-tuning valve is provided at the input port.

[0013] Wherein, the incubator is also provided with a CO 2 Sensors, temperature sensors and humidity sensors.

[0014] Among them, the living cell microscopic imaging analysis device under continuous or intermittent sampling conditions also includes a single filling tube, and the single filling tube is penetrated through the top cover, and the bottom of the single filling tube corresponds to the culture dish.

[0015] The bottom of the single-filling pipe is arranged in a ring-shaped structure, and an oblique spray hole is arranged on the inner side wall of the bottom of the single-filling pipe.

[0016] The beneficial effects of the present invention are:

[0017] The top cover is removed, the culture dish is placed in the culture box, and is fixed firmly by the clamping mechanism. The culture condition of the living cells is analyzed through the microscope. When it is necessary to continuously add samples of the drug, the rotating component is started to drive the rotating ring to rotate on the fixed plate, thereby driving the liquid inlet pipe and the liquid outlet pipe to rotate, so that the ends of the liquid inlet pipe and the liquid outlet pipe located inside the culture box are parallel to the culture dish, and then the pulling component is started to drive the fixed plate to translate, so that the ends of the liquid inlet pipe and the liquid outlet pipe located inside the culture box are located above the culture dish, and then the rotating component is started to drive the liquid inlet pipe and the liquid outlet pipe to rotate through the rotating ring, so that the ends of the liquid inlet pipe and the liquid outlet pipe located inside the culture box are obliquely rotated to the inside of the culture dish, and the continuous addition of the drug is completed by the cooperation of the liquid inlet pipe and the liquid outlet pipe. The above structure can realize continuous addition of the drug, and the versatility is stronger.

[0018] Other advantages, objectives and features of the present invention will be described in the following description to some extent, and to some extent, will be obvious to those skilled in the art based on the following examination and study, or can be taught from the practice of the present invention. The objectives and other advantages of the present invention can be realized and obtained through the following description. BRIEF DESCRIPTION OF THE DRAWINGS

[0019] In order to make the purpose, technical solutions and advantages of the present invention more clear, the present invention will be described in detail below in conjunction with the accompanying drawings, wherein:

[0020] Figure 1 It is a schematic structural diagram of a microscopic imaging analysis device for living cells under continuous or intermittent sample addition conditions according to the first embodiment of the present invention.

[0021] Figure 2 It is a schematic diagram of the external structure of the incubator according to the first embodiment of the present invention.

[0022] Figure 3 The present invention provides Figure 2 A magnified view of the local structure at A.

[0023] Figure 4 It is a schematic diagram of the internal structure of the incubator according to the first embodiment of the present invention.

[0024] Figure 5 The present invention provides Figure 4 A magnified view of the local structure at B.

[0025] Figure 6 It is a schematic structural diagram of a microscopic imaging analysis device for living cells under continuous or intermittent sample addition conditions according to a second embodiment of the present invention.

[0026] Figure 7 Schematic diagram of the structure of a single filling pipe according to the second embodiment of the present invention.

[0027] Figure 8 It is a schematic diagram of the principle of ATP detection in living cells provided by the present invention.

[0028] Fig. 9 The present invention provides a fluorescence response diagram of the ATP concentration change of living cells detected by the microscopic imaging analysis device under the condition of continuous or intermittent sample addition of living cells under the condition of no drug and drug addition.

[0029] 101-incubator, 102-culture dish, 103-top cover, 105-microscope, 106-mounting frame, 107-fixing plate, 108-rotating ring, 109-liquid inlet pipe, 110-liquid outlet pipe, 111-electric telescopic rod, 112-driving motor, 113-output gear, 114-transmission gear, 115-limiting ring, 116-clamping cylinder, 117-arc splint, 118-water tank, 119-temperature regulating module, 120-water inlet, 121-CO 2 Input port, 122-intermittent fine-tuning valve, 123-CO 2 Sensor, 124-temperature sensor, 125-humidity sensor, 201-single filling pipe, 202-oblique spray hole. DETAILED DESCRIPTION

[0030] The present invention is further described below in conjunction with specific implementation methods. The accompanying drawings are only used for exemplary descriptions and are only schematic diagrams, not actual drawings, and cannot be understood as limiting this patent; in order to better illustrate the embodiments of the present invention, some parts of the accompanying drawings may be omitted, enlarged or reduced, and do not represent the size of the actual product; for those skilled in the art, it is understandable that some well-known structures and their descriptions in the accompanying drawings may be omitted.

[0031] Embodiments of the present invention are described in detail below, examples of which are shown in the accompanying drawings, wherein the same or similar reference numerals throughout represent the same or similar elements or elements having the same or similar functions. The embodiments described below with reference to the accompanying drawings are exemplary and are intended to be used to explain the present invention, and should not be construed as limiting the present invention.

[0032] First embodiment:

[0033] See also Figures 1 to 5 ,in Figure 1 is a schematic structural diagram of the microscopic imaging analysis device for living cells under continuous or intermittent sample addition conditions of the first embodiment, Figure 2 is a schematic diagram of the external structure of the incubator of the first embodiment, Figure 3 yes Figure 2 The enlarged view of the local structure at A. Figure 4is a schematic diagram of the internal structure of the incubator of the first embodiment, Figure 5 yes Figure 4 A magnified view of the local structure at B.

[0034] The present invention provides a microscopic imaging analysis device for living cells under continuous or intermittent loading conditions: comprising a culture box 101, a clamping mechanism, a culture dish 102, a top cover 103, a single filling tube 201, a continuous loading mechanism and a microscope 105, wherein the continuous loading mechanism comprises two mounting frames 106, two pulling components, two fixing plates 107, two rotating rings 108, two rotating components, a liquid inlet pipe 109 and a liquid outlet pipe 110. The above scheme solves the problem that the existing living cell microscopic imaging analysis device cannot realize continuous loading of drugs and has poor versatility. It can be understood that the above scheme can be used in the structure of the living cell microscopic imaging analysis device.

[0035] According to this specific embodiment, the culture dish 102 is fixed inside the culture box 101 by the clamping mechanism, a detachable top cover 103 is provided on the top of the culture box 101, and the microscope 105 is installed on the outside of the culture box 101;

[0036] The continuous loading mechanism comprises two mounting frames 106, two pulling assemblies, two fixed plates 107, two rotating rings 108, two rotating assemblies, a liquid inlet pipe 109 and a liquid outlet pipe 110. The two mounting frames 106 are respectively mounted on both sides of the outside of the incubator 101. The fixed plates 107 are mounted on one side of each mounting frame 106 facing the incubator 101 through the pulling assemblies. The rotating ring 108 is rotatably arranged at the center of each fixed plate 107. The two rotating rings 108 are arranged on the other side of the incubator 101. The liquid inlet pipe 109 and the liquid outlet pipe 110 are respectively embedded in the interior, the rotating assembly is arranged on each of the fixed plates 107, the output end of the rotating assembly is mechanically transmitted with the rotating ring 108, the liquid inlet pipe 109 and the liquid outlet pipe 110 both pass through the outer wall of the incubator 101, the liquid inlet pipe 109 and the liquid outlet pipe 110 are both arranged in an L-shaped structure, and one end of the liquid inlet pipe 109 and the liquid outlet pipe 110 located inside the incubator 101 corresponds to the culture dish 102.

[0037] In this embodiment, the top cover 103 is removed, the culture dish 102 is placed in the culture box 101, and is fixed firmly by the clamping mechanism. The culture condition of the living cells is analyzed by the microscope 105. When it is necessary to continuously add drugs, the rotating assembly is started to drive the rotating ring 108 to rotate on the fixed plate 107, thereby driving the liquid inlet pipe 109 and the liquid outlet pipe 110 to rotate, so that one end of the liquid inlet pipe 109 and the liquid outlet pipe 110 located inside the culture box 101 is parallel to the culture dish 102, and then the pulling assembly is started to drive the fixed plate 107 to rotate. After the fixed plate 107 is translated so that one end of the liquid inlet pipe 109 and the liquid outlet pipe 110 located inside the incubator 101 is located above the culture dish 102, the rotating assembly is started, and the liquid inlet pipe 109 and the liquid outlet pipe 110 are driven to rotate through the rotating ring 108, so that the one end of the liquid inlet pipe 109 and the liquid outlet pipe 110 located inside the incubator 101 is obliquely rotated to the inside of the culture dish 102, and the continuous addition of the drug is completed through the cooperation of the liquid inlet pipe 109 and the liquid outlet pipe 110. The above structure can achieve continuous addition of the drug, and is more versatile.

[0038] Furthermore, each of the pulling assemblies is composed of two electric telescopic rods 111 , and the electric telescopic rods 111 are disposed on both sides of each of the fixing plates 107 , and one end of each of the electric telescopic rods 111 away from the fixing plate 107 is connected to the mounting frame 106 .

[0039] In this embodiment, by disposing the electric telescopic rod 111 , the position of the fixing plate 107 can be adjusted, thereby adjusting the position of one end of the liquid inlet pipe 109 and the liquid outlet pipe 110 located inside the incubator 101 .

[0040] Furthermore, each of the rotating components includes a driving motor 112, an output gear 113 and a transmission gear 114. The driving motor 112 is mounted on the fixed plate 107. The output end of the driving motor 112 is provided with the output gear 113. The transmission gear 114 is mounted on the outside of the rotating ring 108, and the output gear 113 is meshed with the transmission gear 114.

[0041] In this embodiment, the driving motor 112 is started to drive the output gear 113 to rotate. Since the output gear 113 is meshed with the transmission gear 114 , the rotating ring 108 is driven to rotate on the fixing plate 107 .

[0042] Furthermore, two limiting rings 115 are disposed outside each rotating ring 108 , and the two limiting rings 115 are respectively located on two sides of the corresponding fixing plate 107 .

[0043] In this embodiment, the limiting ring 115 is provided to make the structure of the rotating ring 108 rotating on the fixing plate 107 more stable.

[0044] Furthermore, the clamping mechanism includes two clamping cylinders 116 and two arc-shaped clamping plates 117 . The clamping cylinders 116 are disposed on both sides of the interior of the incubator 101 . The output end of each clamping cylinder 116 is disposed with the arc-shaped clamping plate 117 , and the arc-shaped clamping plate 117 corresponds to the culture dish 102 .

[0045] In this embodiment, the culture dish 102 can be firmly clamped inside the incubator 101 by the cooperation between the clamping cylinder 116 and the arc-shaped clamping plate 117 .

[0046] Furthermore, the incubator 101 is provided with a water tank 118 and a temperature regulating module 119, wherein the temperature regulating module 119 is used to regulate the ambient temperature in the incubator 101, and the outer wall of the incubator 101 is provided with a water inlet 120 and a CO 2 The water inlet 120 corresponds to the water tank 118. 2 An intermittent fine-tuning valve 122 is provided at the input port 121 .

[0047] In this embodiment, water is poured into the water tank 118 through the water inlet 120, and the temperature in the incubator 101 can be adjusted through the temperature adjustment module 119. The volatilization of water in the water tank 118 is controlled by the change of temperature, thereby adjusting the humidity in the incubator 101. 2 The input port 121 and the intermittent fine-tuning valve 122 can adjust the CO in the incubator 101. 2 content.

[0048] Furthermore, the incubator 101 is also provided with a CO 2 sensor 123 , temperature sensor 124 and humidity sensor 125 .

[0049] In this embodiment, the CO 2 The sensor 123 monitors the CO in the incubator 101 in real time. 2 content, the temperature in the incubator 101 is monitored in real time by the temperature sensor 124, and the humidity in the incubator 101 is monitored in real time by the humidity sensor 125.

[0050] When using the microscopic imaging analysis device for living cells under continuous or intermittent loading conditions of the present embodiment, the top cover 103 is removed, and the culture dish 102 is placed in the culture box 101. The culture dish 102 can be firmly clamped inside the culture box 101 through the cooperation of the clamping cylinder 116 and the arc clamping plate 117. The culture conditions of the living cells are analyzed through the microscope 105. When it is necessary to continuously load the drug, the driving motor 112 is started to drive the output gear 113 to rotate. Since the output gear 113 is meshed with the transmission gear 114, the rotating ring 108 is driven to rotate on the fixed plate 107, thereby driving the liquid inlet pipe 109 and the liquid outlet pipe 110 to rotate, so that the liquid inlet pipe 109 and the liquid outlet pipe 110 are driven to rotate. After one end of the liquid pipe 110 located inside the incubator 101 is parallel to the culture dish 102, the position of the fixed plate 107 can be adjusted by setting the electric telescopic rod 111, so that one end of the liquid inlet pipe 109 and the liquid outlet pipe 110 located inside the incubator 101 is located above the culture dish 102, and then the rotating assembly is started to drive the liquid inlet pipe 109 and the liquid outlet pipe 110 to rotate through the rotating ring 108, so that the ends of the liquid inlet pipe 109 and the liquid outlet pipe 110 located inside the incubator 101 are obliquely rotated to the inside of the culture dish 102, and the continuous addition of the drug is completed by the cooperation of the liquid inlet pipe 109 and the liquid outlet pipe 110. The above structure can realize continuous addition of the drug, and has stronger versatility.

[0051] Second embodiment:

[0052] Based on the first embodiment, please refer to Figure 6 and Figure 7 , Figure 6 This is a schematic diagram of the structure of the microscopic imaging analysis device for living cells under continuous or intermittent sample addition conditions of the second embodiment. Figure 7 It is a schematic structural diagram of a single filling pipe of the second embodiment.

[0053] The invention provides a microscopic imaging analysis device under the condition of continuous or intermittent sample addition of living cells, which also includes a single filling tube.

[0054] According to this specific embodiment, the single filling tube 201 is provided through the top cover 103, and the bottom of the single filling tube 201 corresponds to the culture dish 102. When it is necessary to add medicine intermittently to the culture dish 102, the intermittent addition of medicine can be completed through the setting of the single filling tube 201, and the operation is simple.

[0055] Among them, the bottom of the single filling pipe 201 is arranged in an annular structure, and an oblique spray hole 202 is arranged on the inner side wall of the bottom of the single filling pipe 201. Since the bottom of the single filling pipe 201 is arranged in an annular structure, and the oblique spray hole 202 is arranged on the inner side wall of the bottom of the single filling pipe 201, it is smoother to add samples into the culture dish 102 through the single filling pipe 201.

[0056] When using a microscopic imaging analysis device for living cells under continuous or intermittent loading conditions of the present embodiment, when it is necessary to intermittently add drugs to the culture dish 102, the intermittent loading of drugs can be completed through the setting of the single filling tube 201, and the operation is simple. In addition, since the bottom of the single filling tube 201 is arranged in an annular structure, and the oblique spray hole 202 is arranged on the inner side wall of the bottom of the single filling tube 201, the loading of samples into the culture dish 102 through the single filling tube 201 is smoother.

[0057] Application Example: The application of the present invention in monitoring ATP expression in living cells under drug administration conditions is described in detail below.

[0058] Among them, the principle of living cell ATP detection is as follows Figure 8 As shown: In the absence of ATP, the nucleic acid probe is in an open structure "OFF State" and there is no fluorescent molecule binding domain; in the presence of ATP, the target molecule specifically interacts with the nucleic acid probe, inducing a structural change in the nucleic acid probe, prompting the three-dimensional structure of the nucleic acid probe to change to a closed structure "ON State", forming a fluorescent molecule binding domain, and then specifically identifying the fluorescent molecule to generate a fluorescent signal; since the closed structure of the nucleic acid probe is stably present under the synergistic effect of the target molecule and the fluorescent molecule, when the target molecule is dissociated / degraded, the nucleic acid probe gradually changes from a closed structure "ON State" to an open structure "OFF State", and the fluorescent molecule dissociates from the nucleic acid probe, and the fluorescent signal decreases. Based on this, the structural change of the nucleic acid probe is mediated by ATP, resulting in changes in the interaction between the fluorescent molecule and the nucleic acid probe, and then the change in ATP molecule concentration is monitored through the dose-effect relationship between the fluorescence intensity and the ATP concentration.

[0059] The specific application is: pre-mix the nucleic acid probe that forms the fluorescent molecule binding domain with the transfection reagent to form a detection solution. Add 500 μL of the detection model cell Hela cell to the culture dish 102, pre-culture for 4 hours, and after it is completely attached to the wall, use phosphate buffer solution and culture medium to fully wash, add culture medium and detection solution, and place it in the incubator 101 involved in the technical solution, set the humidity and carbon dioxide concentration; after 2 hours of pre-transfection, connect the drug to be added to the single filling tube 201 or the continuous sample addition mechanism; set the fluorescence detection conditions, including excitation wavelength, detection wavelength, sampling interval time, exposure time, drug administration time, detection time, etc., and use the detection method to detect the change of ATP concentration of living cells under drug-free and drug-added conditions.

[0060] like Fig. 9 As shown, compared with Hela cells without drug effect ( Fig. 9 A), when Hela cells are treated with calcium ions, their fluorescence response signal is significantly reduced, indicating a lower ATP content level; the drug administration path of the microscopic imaging analysis device under the continuous or intermittent loading conditions of living cells is switched (the drug to be added is connected to the single filling tube 201 or the continuous loading mechanism), and the oligomycin drug action is cut in, and its fluorescence response signal is significantly enhanced, indicating that the ATP content level is enhanced. The results show that the microscopic imaging analysis device under the continuous or intermittent loading conditions of living cells involved in the technical solution can be used for long-term monitoring of living cell-related molecules under drug administration conditions.

[0061] Finally, it should be noted that the above embodiments are only used to illustrate the technical solution of the present invention rather than to limit it. Although the present invention has been described in detail with reference to the preferred embodiments, those skilled in the art should understand that the technical solution of the present invention can be modified or replaced by equivalents without departing from the purpose and scope of the technical solution, which should be included in the scope of the claims of the present invention.

Claims

1. A microscopic imaging analysis device for living cells under continuous or intermittent sample addition conditions, characterized in that: The device comprises an incubator, a clamping mechanism, a culture dish, a top cover, a continuous sample loading mechanism and a microscope, wherein the culture dish is fixed inside the incubator through the clamping mechanism, a detachable top cover is arranged on the top of the incubator, and the microscope is installed on the outside of the incubator; The continuous loading mechanism includes two mounting frames, two pulling components, two fixed plates, two rotating rings, two rotating components, a liquid inlet pipe and a liquid outlet pipe. The two mounting frames are respectively mounted on both sides of the outside of the incubator, and the fixed plate is mounted on one side of each mounting frame facing the incubator through the pulling component. The rotating ring is rotatably arranged at the center of each fixed plate, and the liquid inlet pipe and the liquid outlet pipe are respectively embedded in the interiors of the two rotating rings. The rotating component is arranged on each fixed plate, and the output end of the rotating component is mechanically transmitted with the rotating ring. The liquid inlet pipe and the liquid outlet pipe both pass through the outer wall of the incubator, and the liquid inlet pipe and the liquid outlet pipe are both arranged in an L-shaped structure. The ends of the liquid inlet pipe and the liquid outlet pipe located inside the incubator correspond to the culture dish.

2. The living cell microscopic imaging analysis device under continuous or intermittent sample addition conditions as claimed in claim 1, characterized in that: Each of the pulling components is composed of two electric telescopic rods, and the electric telescopic rods are arranged on both sides of each of the fixing plates, and one end of each of the electric telescopic rods away from the fixing plate is connected to the mounting frame.

3. The living cell microscopic imaging analysis device under continuous or intermittent sample addition conditions as claimed in claim 2, characterized in that: Each of the rotating components includes a driving motor, an output gear and a transmission gear. The driving motor is mounted on the fixed plate. The output end of the driving motor is provided with the output gear. The transmission gear is mounted on the outside of the rotating ring, and the output gear is meshed with the transmission gear.

4. The living cell microscopic imaging analysis device under continuous or intermittent sample addition conditions as claimed in claim 3, characterized in that: Two limiting rings are arranged outside each rotating ring, and the two limiting rings are respectively located on two sides of the corresponding fixing plate.

5. The living cell microscopic imaging analysis device under continuous or intermittent sample addition conditions as claimed in claim 4, characterized in that: The clamping mechanism comprises two clamping cylinders and two arc-shaped clamping plates. The clamping cylinders are arranged on both sides of the interior of the incubator. The output end of each clamping cylinder is arranged with the arc-shaped clamping plate, and the arc-shaped clamping plate corresponds to the culture dish.

6. The living cell microscopic imaging analysis device under continuous or intermittent sample addition conditions as claimed in claim 5, characterized in that: A water tank and a temperature regulating module are also provided inside the incubator. The temperature regulating module is used to adjust the ambient temperature inside the incubator. A water inlet and a CO2 inlet are also provided on the outer wall of the incubator. The water inlet corresponds to the water tank. An intermittent fine-tuning valve is provided at the CO2 inlet.

7. The living cell microscopic imaging analysis device under continuous or intermittent sample addition conditions as claimed in claim 6, characterized in that: The incubator is also provided with a CO2 sensor, a temperature sensor and a humidity sensor.

8. The living cell microscopic imaging analysis device under continuous or intermittent sample addition conditions as claimed in claim 1, characterized in that: The living cell microscopic imaging analysis device under continuous or intermittent sample addition conditions further comprises a single filling tube, which is penetrated through the top cover and has a bottom corresponding to the culture dish.

9. The living cell microscopic imaging analysis device under continuous or intermittent sample addition conditions as claimed in claim 8, characterized in that: The bottom of the single-filling pipe is arranged in an annular structure, and an oblique spray hole is arranged on the inner side wall of the bottom of the single-filling pipe.

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