Multifunctional cell ball culture observation device

Abstract

The utility model relates to the technical field of cell culture accessories, in particular to a multifunctional cell ball culture observation device which comprises a culture dish, a supporting device, a reflecting mirror and a microscope, and the supporting device comprises an out-dish control handle and an in-dish mirror surface fixing piece; a reflector is clamped by adopting an in-dish mirror surface fixing piece, and the in-dish mirror surface fixing piece and an out-dish control handle are arranged on the two sides of a culture dish cover and are fixed through magnets; when the side surface of the sample is shot, a side-shooting light source is provided for the bottom of the culture dish, and an image of the sample is reflected to the microscope through the reflector; the focal length of the objective lens of the microscope is adjusted to display a clear image of the sample in the reflector, the side surface of the sample is obtained, the observation device does not interfere with the growth process of the sample, and sample pollution is reduced to the greatest extent; and multi-angle observation and shooting of three-dimensional cells can be carried out through a conventional microscope.

Description

Technical Field

[0001] The utility model relates to the technical field of cell culture accessories, in particular to a multifunctional cell sphere culture observation device and an observation and shooting method. Background Art

[0002] Three-dimensional (3D) cell culture is an experimental technique that simulates the in-vivo cell microenvironment. It allows cells to grow and differentiate in three-dimensional space. Compared with two-dimensional (2D) cell culture, 3D culture is closer to the physiological state of a real organism. In 3D culture, cells can form complex structures such as spheroids and organoids, which are of great value in studying cell behavior, drug screening, organ development, and disease models. Among them, the 3D tumor sphere cell model has become a powerful tool for studying tumor biology and drug response. Compared with traditional two-dimensional cell culture models, these multicellular aggregates better reproduce the complex microenvironment and heterogeneity of solid tumors, exhibit gradients of oxygen, nutrients, and metabolites, and simulate the pathophysiological processes of primary tumors. Therefore, they are of great value in studying tumor growth, invasion, and response to treatment. Organoids are more complex three-dimensional structure miniature tissues cultured in vitro. They not only contain multiple types of cells but also simulate a part of the corresponding organ or tissue in terms of structure and function. Organoids can be formed from stem cells or mature cells through a self-organization process to form 3D cell spheres, and they show great potential in simulating organ development, disease model construction, and personalized medicine.

[0003] Characterizing the morphology and volume of three-dimensional (3D) growing cells is crucial for understanding their growth kinetics and evaluating treatment effects. Typically, it is assumed that 3D growing cells are perfect spheres or have centrosymmetric or axisymmetric morphologies. Researchers usually estimate the volume by measuring the bottom diameter. However, this method is clearly not applicable to cell models with complex 3D structures of spheres and has low generality, and can only be used to measure 3D cell models with relatively simple shapes. More accurate volume calculations can be performed by directly measuring the thickness, which can better restore the non-uniform morphology and true size of 3D cells. For the growth of 3D cell spheres, their length, width, and height changes are basically logarithmic growth. Once the sphere exceeds the critical diameter (usually >500 microns), it enters the steady state of volume growth. At this point, oxygen and nutrients diffuse into the core of the sphere, greatly limiting the growth of the 3D cell sphere, and the 3D cell sphere cannot grow indefinitely. If the size changes of 3D cells in the z-direction height can be further measured, it can provide a more sensitive growth arrest reading than the lengths in the x- and y-directions. In addition to monitoring the growth of 3D cell spheres, the migration of 3D cell spheres and the fusion process between 3D cell spheres are also difficult to observe non-invasively and in real-time in situ by conventional observation methods. If the embedding and staining observations are carried out after the migration and fusion of 3D cells are completed, it will greatly affect the original structure of the cell sphere and cannot observe the specific change process.

[0004] Currently, the optical microscopes used in the laboratory have great limitations in quantifying the complex 3D structures of spheres. When measuring the stereoscopic morphology of 3D cell spheres based on two dimensions, the mutual occlusion of semi-transparent cells hinders the measurement of the image thickness in the z-direction. Confocal imaging can optically section the sphere, generate 3D reconstruction images, and by fluorescently labeling the cells, the thickness and diameter can be directly measured from the cross-sectional view. However, light scattering and absorption limit the imaging depth, making it challenging to resolve the core of large spheres (>300 microns).

[0005] Optical coherence tomography (OCT) is a non-invasive imaging technique that uses low-coherence light to create cross-sectional images of tissue structures and can measure the thickness and diameter even in large spheres. Light-sheet fluorescence microscopy (LSFM) is another emerging technology that uses a thin light sheet to illuminate the sample, can quickly perform 3D imaging of the entire sphere, and measure the thickness and diameter through 3D reconstruction. However, LSFM requires specialized equipment and may be limited by light scattering in the dense sphere core. For the above-mentioned 3D imaging methods of the stereostructure of 3D cells, it is very difficult to monitor the real-time in situ 3D structural changes of 3D cells. Summary of the Utility Model

[0006] The purpose of the present utility model is to provide a multifunctional cell sphere culture observation device and an observation and shooting method, aiming to solve the problem that currently in the laboratory, three-dimensional cell sphere culture cannot be carried out in a culture dish while observing the morphological and structural changes of the cell sphere without disturbance.

[0007] To achieve the above object, in the first aspect, the present utility model provides a multifunctional cell sphere culture observation device, including a culture dish, a support device, a reflector and a microscope. The support device includes an external dish control handle and an internal dish mirror fixing member.

[0008] The microscope is arranged below the culture dish, the external dish control handle is arranged on the top of the culture dish, the internal dish mirror fixing member is arranged inside the culture dish, the reflector is clamped with the internal dish mirror fixing member and is located on the side of the internal dish mirror fixing member away from the external dish control handle.

[0009] Among them, the internal dish mirror fixing member has a notch, and the notch is located on the side away from the external dish control handle.

[0010] Among them, the reflector is fixed in the notch and maintains an angle of 45° with the horizontal plane.

[0011] Among them, the culture dish includes a culture dish bottom and a culture dish cover. The culture dish bottom is arranged above the microscope, and the culture dish cover is clamped with the culture dish bottom and is located on the top of the culture dish bottom.

[0012] In the second aspect, the present utility model also provides a multifunctional cell sphere culture observation device observation and shooting method, including the following steps:

[0013] Clamp the reflector with the internal dish mirror fixing member, and place the internal dish mirror fixing member and the external dish control handle on both sides of the culture dish cover and fix them with magnets.

[0014] When photographing the side of the sample, provide a side shooting light source to the culture dish bottom, and the image of the side of the sample is reflected by the reflector to the microscope.

[0015] Adjust the focal length of the microscope objective lens so that a clear image of the sample is displayed in the reflector to obtain the image of the side of the sample.

[0016] A multifunctional cell sphere culture observation device of the present utility model clamps the mirror using the in-dish mirror fixing member, places the in-dish mirror fixing member and the out-dish control handle on both sides of the culture dish cover, and fixes them with magnets. When photographing the side of the sample, a side-shot light source (non-microscope light source) is provided to the bottom of the culture dish, and the image of the sample is reflected by the mirror to the microscope. The focal length of the microscope objective is adjusted so that a clear image of the sample is shown in the mirror, and an image of the side of the sample is obtained. This observation can perform multi-angle observation and photographing of three-dimensional cells through a conventional laboratory microscope without expensive facilities and equipment, achieving non-invasive in-situ monitoring of the side of three-dimensional cell spheres (since the support device and the mirror are both placed in the culture environment and do not need to be repeatedly taken, there will be no displacement of the sample and the photographing device); it realizes that the device does not need to repeatedly open and close the culture dish cover when photographing the side of three-dimensional cell spheres, greatly reducing its contamination probability (once the support device and the mirror are installed, the culture dish cover does not need to be opened); it greatly reduces the volume of the multifunctional cell sphere culture and multi-angle observation device, and can be adapted to a smaller cell monitoring platform and a narrow environment with more position restrictions for three-dimensional cell side observation and photographing. It solves the problem that currently in the laboratory, it is impossible to perform non-invasive observation of the morphological structure changes of cell spheres while culturing three-dimensional cell spheres in a culture dish. BRIEF DESCRIPTION OF THE DRAWINGS

[0017] In order to more clearly illustrate the technical solutions in the embodiments of the present utility model or the prior art, the following will briefly introduce the drawings required for use in the description of the embodiments or the prior art. Obviously, the following drawings are only some embodiments of the present utility model. For those of ordinary skill in the art, without creative efforts, other drawings can be obtained based on these drawings.

[0018] Figure 1 It is a schematic diagram of a multifunctional cell sphere culture observation device provided by the present utility model.

[0019] Figure 2 It is a structural schematic diagram of a multifunctional cell sphere culture observation device provided by the present utility model.

[0020] Figure 3 It is a schematic diagram of the use of a multifunctional cell sphere culture observation device provided by the present utility model.

[0021] Figure 4 It is a schematic diagram of taking a photo in Embodiment 1.

[0022] Figure 5 It is a schematic diagram of taking a photo in Embodiment 2.

[0023] Figure 6 It is a schematic diagram of taking a photo in Embodiment 3.

[0024] Figure 7 It is a schematic side view of the structure of the ultra-low adhesion microporous culture array side shooting device in Example 4.

[0025] Figure 8 It is a schematic diagram of the cooperation between the mirror and the cell sphere culture array.

[0026] Figure 9 It is a schematic diagram of the use of the ultra-low adhesion microporous culture array side shooting device.

[0027] Figure 10 Flowchart of the observation and shooting method of a multifunctional cell sphere culture observation device provided by the present utility model.

[0028] In the figure: 1 - mirror, 2 - microscope, 3 - outer dish control handle, 4 - inner dish mirror fixing part, 5 - notch, 6 - culture dish bottom, 7 - culture dish cover. Specific embodiments

[0029] The embodiments of the present utility model will be described in detail below. The examples of the embodiments are shown in the accompanying drawings, where the same or similar reference numerals represent the same or similar elements or elements with the same or similar functions from beginning to end. The embodiments described below with reference to the accompanying drawings are exemplary and are intended to explain the present utility model, and should not be construed as a limitation of the present utility model.

[0030] Please refer to Figures 1 to 9 , in the first aspect, the present utility model provides a multifunctional cell sphere culture observation device, including a culture dish, a support device, a mirror 1 and a microscope 2. The support device includes an outer dish control handle 3 and an inner dish mirror fixing part 4; the microscope 2 is arranged below the culture dish, the outer dish control handle 3 is arranged on the top of the culture dish, the inner dish mirror fixing part 4 is arranged inside the culture dish, the mirror 1 is clamped with the inner dish mirror fixing part 4 and is located on the side of the inner dish mirror fixing part 4 away from the outer dish control handle 3; the inner dish mirror fixing part 4 has a notch 5, the notch 5 is located on the side away from the outer dish control handle 3, the culture dish includes a culture dish bottom 6 and a culture dish cover 7, the culture dish bottom 6 is arranged above the microscope 2, and the culture dish cover 7 is clamped with the culture dish bottom 6 and is located on the top of the culture dish bottom 6.

[0031] In this embodiment, the in-dish mirror fixing member 4 is used to clamp the mirror 1, and the in-dish mirror fixing member 4 and the out-dish control handle 3 are placed on both sides of the culture dish cover 7 and fixed by magnets. When photographing the side of the sample, a side shooting light source (non-microscope light source) is provided to the culture dish bottom 6, and the image of the sample is reflected by the mirror 1 to the microscope 2. The objective lens focal length of the microscope 2 is adjusted so that a clear image of the sample is shown in the mirror 1, and the side of the sample is obtained. This observation can be carried out on three-dimensional cells from multiple angles through a conventional laboratory microscope 2 without expensive facilities and equipment, realizing non-invasive in-situ monitoring of the side of the three-dimensional cell sphere (since the support device and the mirror 1 are both placed in the culture environment and do not need to be repeatedly taken, which may cause displacement of the sample and the photographing device); it is realized that the device does not need to repeatedly open and close the culture dish cover when photographing the side of the three-dimensional cell sphere, greatly reducing the probability of its contamination (once the support device and the mirror 1 are installed, it may not be necessary to open the culture dish cover 7); the volume of the multi-functional cell sphere culture and multi-angle observation device is greatly reduced, and it can be adapted to a smaller cell monitoring platform and a narrow environment with more position restrictions for observing and photographing the side of three-dimensional cells. It solves the problem that currently in the laboratory, three-dimensional cell spheres cannot be cultured in a culture dish while observing the morphological and structural changes of the cell spheres without disturbance.

[0032] Among them, the support device further includes a coated mirror, which is used in combination with an ultra-low adhesion microporous array culture dish. The coated mirror is located on the side surface of the support device. The ultra-low adhesion microporous array culture dish is integrally formed and is used for directly carrying out the spheroid formation experiment of three-dimensional cells. The support device is placed at the central void position of the ultra-low adhesion microporous array culture dish. Through the reflection of the coated mirror 1 on the side surface of the support device, the side morphological changes of the three-dimensional cells are reflected by the coated mirror, and then the microscope 2 is used to observe the image in the coated mirror to take pictures of the side image of the three-dimensional cell sample.

[0033] Please refer to Figure 10 , on the second aspect, the present invention also provides a method for observing and photographing a multi-functional cell sphere culture and observation device, including the following steps:

[0034] S1 Use the in-dish mirror fixing member 4 to clamp the mirror 1, and place the in-dish mirror fixing member 4 and the out-dish control handle 3 on both sides of the culture dish cover 7 and fix them by magnets;

[0035] S2 When photographing the side of the sample, provide a side shooting light source to the culture dish bottom 6, and the side image of the sample is reflected by the mirror 1 to the microscope 2;

[0036] S3 Adjust the objective lens focal length of the microscope 2 so that a clear image of the sample is shown in the mirror 1 to obtain the side image of the sample.

[0037] In order to better understand the present technical solution, the following examples are provided for further explanation:

[0038] Example 1

[0039] Step 1: Use the inverted microscope 2 with the multifunctional spheroid culture and multi-angle observation device of the first embodiment to observe the bottom view and side view of the three-dimensional spheroid formation process.

[0040] Step 2: Place the cell culture agarose array in a cell culture dish and add 1×10 4 Number of human prostate cancer cells, and 2 mL of complete culture medium was added around the array to maintain cell growth;

[0041] Step 3: Magnetically connect the side camera device to the cell culture dish cover 7. After closing the dish cover, adjust the support device so that the reflector 1 is parallel to the cell culture agarose array. There is no need to adjust the position of the reflector 1 later.

[0042] Step 4: Turn on microscope 2 and its built-in light source, place the cell culture dish containing the cell culture agarose array on the stage of inverted microscope 2, adjust the light source to an appropriate brightness, observe the state of the sample in the eyepiece of microscope 2, find the image of the bottom surface of the sample in a larger field of view, and adjust the coarse and fine focusing knobs of microscope 2 in sequence until a clear image appears in the field of view;

[0043] Step 5: Switch different objective lenses to take multiple magnification images of the sample bottom surface;

[0044] Step 6: Place the external side-mounted mercury lamp on the side of the cell culture dish facing the reflector 1 and adjust it to a suitable brightness;

[0045] Step 7. Switch back to a larger field of view, find the image of the sample in the reflector in the eyepiece, and adjust the coarse and fine focus knobs in sequence until the image in the field of view is clear;

[0046] Step 8: Switch different objective lenses to take multiple side images of the sample.

[0047] Step 9. Place the cells in a carbon dioxide incubator and take them out and take pictures every 12 hours.

[0048] Example 2

[0049] Step 1: Use the inverted microscope 2 in conjunction with the multifunctional spheroid culture and multi-angle observation device of the first embodiment to observe the bottom view and side view of the three-dimensional spheroid migration process.

[0050] Step 2: The initial number of cells cultured in agarose array for 7 days was 1×10 4 Transfer the human prostate cancer cell spheres into a cell culture dish and add 2 mL of complete culture medium into the cell culture dish;

[0051] Step 3: Magnetically connect the side camera device to the cell culture dish cover 7. After the dish cover is closed, adjust the support device under the microscope 2 so that the reflector 1 is close to the cell sphere placed in the cell culture dish. There is no need to adjust the position of the reflector 1 later.

[0052] Step 4: Turn on microscope 2 and its built-in light source, place the cell culture dish containing the cell culture agarose array on the stage of inverted microscope 2, adjust the light source to an appropriate brightness, observe the state of the sample in the eyepiece of microscope 2, find the image of the bottom surface of the sample in a larger field of view, and adjust the coarse and fine focusing knobs of microscope 2 in sequence until a clear image appears in the field of view;

[0053] Step 5: Switch different objective lenses to take multiple magnification images of the sample bottom surface;

[0054] Step 6: Place the external side-mounted mercury lamp on the side of the cell culture dish facing the reflector 1 and adjust it to a suitable brightness;

[0055] Step 7. Switch back to a larger field of view, find the image of the sample in the reflector in the eyepiece, and adjust the coarse and fine focus knobs in sequence until the image in the field of view is clear;

[0056] Step 8: Switch different objective lenses to take multiple side images of the sample.

[0057] Step 9. Place the cells in a 37°C carbon dioxide incubator and take them out and photograph them every 12 hours.

[0058] Example 3

[0059] Step 1: Use the inverted microscope 2 in conjunction with the multifunctional spheroid culture and multi-angle observation device of the first embodiment to observe the bottom view and side view of the three-dimensional spheroid fusion process.

[0060] Step 2: Place the cell culture agarose array in a cell culture dish and add 7-day-old cells to each well of the array with an initial number of 1×10 4 Human prostate cancer cell spheres were transferred to one well of the agarose array, and 2 mL of complete culture medium was added around the array to maintain cell growth.

[0061] Step 3: Magnetically connect the side camera device to the cell culture dish cover 7. After closing the dish cover, adjust the support device so that the reflector 1 is parallel to the cell culture agarose array. There is no need to adjust the position of the reflector 1 later.

[0062] Step 4: Turn on microscope 2 and its built-in light source. Place the cell culture dish containing the cell culture agarose array on the stage of the inverted microscope 2. Adjust the light source to an appropriate brightness, observe the state of the sample in the eyepiece of microscope 2, find the image of the bottom surface of the sample in a larger field of view, and sequentially adjust the coarse focus screw and fine focus screw of microscope 2 until a clear image appears in the field of view;

[0063] Step 5: Switch to different objective lenses and take pictures of the image of the bottom surface of the sample at multiple magnifications;

[0064] Step 6: Place the external side-shot mercury lamp on the side of the cell culture dish facing mirror 1 and adjust it to an appropriate brightness;

[0065] Step 7: Switch back to a larger field of view, find the image of the sample in the reflector in the eyepiece, and sequentially adjust the coarse focus screw and fine focus screw until the image in the field of view is clear;

[0066] Step 8: Switch to different objective lenses and take pictures of the image of the side surface of the sample at multiple magnifications,

[0067] Step 9: Place the cells in a 37°C carbon dioxide incubator and take pictures every 12 hours.

[0068] Example of using with an ultra-low adhesion microporous array culture dish:

[0069] The whole device is divided into three parts: a light source cover, an ultra-low adhesion annular cell sphere culture dish, and a reflector assembly. Among them, the ultra-low adhesion annular cell sphere culture dish is divided into a culture array upper cover and an ultra-low adhesion annular cell sphere culture array. The ultra-low adhesion annular cell sphere culture array is made of a low adhesion substrate, and cells can form spheres in the array holes. The bottom of the reflector assembly is inclined at a 45-degree angle, and the surface is coated with a mirror coating to achieve the same effect as a mirror. There is a surface light source on the top of the light source cover, and an annular light source is wound around the bottom of the side wall, and the brightness can be adjusted.

[0070] Example 4

[0071] Step 1: Use an inverted microscope to observe the bottom view and side view of the three-dimensional cell sphere formation process of the ultra-low adhesion microporous culture array side-shot device in the second embodiment.

[0072] Step 2: Add 1×10 4 number of human prostate cancer cells into each micropore of the ultra-low adhesion array, and add 3 mL of complete medium around the array to maintain the cell growth state;

[0073] Step 3: Place the mirror assembly at the central cavity position of the ultra-low adhesion array. After placing it stably, the mirror should be flush with the top surface of the ultra-low adhesion array. Cover the upper cover of the culture array, and there is no need to adjust the position of the mirror subsequently.

[0074] Step 4: When taking the bottom and side images of the ultra-low adhesion array, place the culture dish of the ultra-low adhesion array containing cell spheres on the stage of the inverted microscope 2. Place the light source cover to cover the ultra-low adhesion cell culture array, and observe in the eyepiece. After adjusting the light source to an appropriate brightness, check the sample status.

[0075] Step 5: Locate the image of the bottom surface of the sample in a larger field of view in the eyepiece, and sequentially adjust the coarse focusing screw and the fine focusing screw of the microscope 2 until a clear image appears in the field of view.

[0076] Step 6: Switch to different objective lenses to take images of the bottom surface of the sample at multiple magnifications.

[0077] Step 7: Switch back to a larger field of view, locate the image of the sample in the mirror assembly in the eyepiece, and sequentially adjust the coarse focusing screw and the fine focusing screw until the image in the field of view is clear.

[0078] Step 8: Switch to different objective lenses to take images of the side surface of the sample at multiple magnifications.

[0079] Step 9: Place the cells in a carbon dioxide incubator and take pictures every 12 hours.

[0080] Example 5

[0081] Step 1: Use the inverted microscope 2 in combination with the side shooting device of the ultra-low adhesion micro-well culture array in the second embodiment to observe the bottom view and side view of the killing process of three-dimensional cell spheres of doxorubicin (DOX).

[0082] Step 2: Place one three-dimensional cell sphere of personal prostate cancer cells that has grown for 7 days and has an initial number of 1×10 4 into each micro-well of the ultra-low adhesion array, and add 3 mL of complete medium containing 50 μg / mL DOX around the array to maintain the cell growth state and observe the drug killing of the cell spheres.

[0083] Step 3: Place the mirror assembly at the central cavity position of the ultra-low adhesion array. After placing it stably, the mirror should be flush with the top surface of the ultra-low adhesion array. Cover the upper cover of the culture array, and there is no need to adjust the position of the mirror subsequently.

[0084] Step 4: When taking the bottom and side images of the ultra-low adhesion array, place the cell culture dish of the ultra-low adhesion cell culture array on the stage of the inverted microscope 2. Place the light source hood to cover the ultra-low adhesion cell culture array. Observe through the eyepiece, adjust the light source to an appropriate brightness, and then check the sample status.

[0085] Step 5: Find the image of the bottom surface of the sample in a larger field of view through the eyepiece. Adjust the coarse focusing screw and the fine focusing screw of the microscope 2 in sequence until a clear image appears in the field of view.

[0086] Step 6: Switch to different objective lenses to take images of the bottom surface of the sample at multiple magnifications.

[0087] Step 7: Switch back to a larger field of view. Find the image of the sample in the mirror assembly through the eyepiece. Adjust the coarse focusing screw and the fine focusing screw in sequence until the image in the field of view is clear.

[0088] Step 8: Switch to different objective lenses to take images of the side surface of the sample at multiple magnifications.

[0089] Step 9: Place the cells in a carbon dioxide incubator and take pictures every 12 hours.

[0090] Example 6

[0091] Use the inverted microscope 2 in combination with the side shooting device of the ultra-low adhesion microporous culture array in the second example to observe the bottom view and side view of the process of three-dimensional cell spheroid natural killer cells (NK cells).

[0092] Step 1: Add 1×10 4 number of human prostate cancer cells to each micropore of the ultra-low adhesion array, and add 1×10 4 number of NK cells to each micropore of the array;

[0093] Step 2: Add 3 mL of special medium for NK cells around the array to maintain the cell growth state.

[0094] Step 3: Place the mirror assembly at the central cavity position of the ultra-low adhesion array. After placing it stably, the mirror should be flush with the top surface of the ultra-low adhesion array. Cover the upper cover of the culture array, and there is no need to adjust the position of the mirror subsequently.

[0095] Step 4: When taking the bottom and side images of the ultra-low adhesion array, place the cell culture dish containing the cell culture agarose array on the stage of the inverted microscope 2. Place the light source hood to cover the ultra-low adhesion cell culture array. Observe through the eyepiece, adjust the light source to an appropriate brightness, and then check the sample status.

[0096] Step 5: Locate the image of the bottom surface of the sample in a larger field of view in the eyepiece, and sequentially adjust the coarse focusing screw and the fine focusing screw of the microscope 2 until a clear image appears in the field of view;

[0097] Step 6: Switch to different objective lenses and take images of the bottom surface of the sample at multiple magnifications;

[0098] Step 7: Switch back to the larger field of view, locate the image of the sample in the mirror assembly in the eyepiece, and sequentially adjust the coarse focusing screw and the fine focusing screw until the image in the field of view is clear;

[0099] Step 8: Switch to different objective lenses and take images of the side surface of the sample at multiple magnifications;

[0100] Step 9: Place the cells in a 37°C carbon dioxide incubator and take pictures every 12 hours after taking them out.

[0101] The above-disclosed is only a preferred embodiment of a multifunctional cell sphere culture observation device and an observation and shooting method of the present utility model. Of course, the scope of the rights of the present utility model cannot be limited thereby. Those of ordinary skill in the art can understand all or part of the processes of implementing the above embodiments, and the equivalent changes made according to the claims of the present utility model still fall within the scope covered by the utility model.

Claims

1. A multifunctional cell sphere culture observation device, characterized in that it includes a culture dish, a support device, a mirror and a microscope, and the support device includes an external dish control handle and an internal dish mirror fixing member; the microscope is arranged below the culture dish, the external dish control handle is arranged on the top of the culture dish, the internal dish mirror fixing member is arranged inside the culture dish, the mirror is clamped with the internal dish mirror fixing member and is located on the side of the internal dish mirror fixing member away from the external dish control handle.

2. The multifunctional cell sphere culture observation device according to claim 1, characterized in that the internal dish mirror fixing member has a notch, and the notch is located on the side away from the external dish control handle.

3. The multifunctional cell sphere culture observation device according to claim 2, characterized in that the mirror is fixed in the notch and maintains an angle of 45° with the horizontal plane.

4. The multifunctional cell sphere culture observation device according to claim 1, characterized in that the culture dish includes a culture dish bottom and a culture dish cover, the culture dish bottom is arranged above the microscope, the culture dish cover is clamped with the culture dish bottom and is located on the top of the culture dish bottom.

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