Multi-angle observation device with multifunctional cell balling culture dish
By designing a multi-angle observation device with a multifunctional cell spheroid culture dish, and using mirror and microscope technology, three-dimensional side observation of cells was realized. This solved the problem of difficulty in observing the morphological changes of the side of cell spheroids in situ in existing technologies, and reduced the risk of contamination and operational difficulty.
Patent Information
- Application Number
- CN202520105418.2
- Authority / Receiving Office
- CN · China
- Patent Type
- Utility models(China)
- Current Assignee / Owner
- Filing Date
- 2025-01-16
- Publication Date
- 2026-01-27
- Estimated Expiration
- 2035-01-16
AI Technical Summary
Existing three-dimensional cell culture technology makes it difficult to observe changes in the lateral morphology of cell spheres in situ while maintaining stable growth. Furthermore, conventional imaging methods are difficult to achieve real-time, non-contact dynamic observation, and the operation is cumbersome and has an impact on cell spheres and the environment.
Design a multi-angle observation device with a multifunctional cell spheroidization culture dish, including a base, shell cover, battery pack, light strip, control board, support device, reflector and three-dimensional cell culture array. The reflector reflects images of different sides of the cells, and combined with a microscope, multi-angle imaging is performed to ensure that the cell growth state remains unchanged and reduce the risk of contamination.
It enables lateral observation of three-dimensional cells, reduces the risk of contamination, simplifies operation, and allows even those without professional training to perform lateral imaging of three-dimensional cells, providing an efficient and economical solution.
Smart Images
Figure CN223837438U_ABST
Abstract
Description
Technical Field
[0001] This utility model relates to the field of cell culture accessories technology, and in particular to a multi-angle observation device with a multifunctional cell spheroidization culture dish. Background Technology
[0002] In biomedical research, three-dimensional (3D) cell culture technology is gradually emerging, especially in the fields of cell biology and oncology. Compared to traditional two-dimensional (2D) culture, 3D models can more accurately simulate the complex microenvironment and heterogeneity of solid tumors, which is of great significance for understanding tumor growth, invasion, and drug response. However, compared to the complete and functionally defined characterization methods of traditional two-dimensional cell systems, the characterization of three-dimensional cell spheres, such as organoids, still faces many challenges.
[0003] Traditional three-dimensional culture methods and optical microscopy have limitations in the quantitative analysis of 3D cell spheroids, as two-dimensional images cannot accurately reflect their three-dimensional characteristics, especially for structurally complex cell spheroids. Therefore, more advanced technologies are needed to accurately measure the thickness of cell spheroids in the Z-axis to obtain their volume and structural properties.
[0004] Furthermore, the growth, migration, and fusion processes of cell spheroids in three-dimensional space are highly complex and dynamic, making real-time, non-contact dynamic observation difficult with conventional imaging methods. While existing technologies such as confocal microscopy provide a certain level of visualization, they are still limited by light scattering and imaging depth, making it difficult to resolve information in the core region of larger spheroids and maintain the cell spheroid's growth state during observation. Additionally, the side-viewing platform for cell spheroids requires extremely high sterility in its device design, is cumbersome to operate, and demands high technical skills from the experimenters, increasing the experimental difficulty and potentially causing unnecessary impacts on the cell spheroids and their surrounding environment. Utility Model Content
[0005] The purpose of this invention is to provide a multi-angle observation device with a multifunctional cell spheroid culture dish, which aims to solve the problem that existing three-dimensional cell spheroids cannot observe lateral morphological changes in situ while maintaining a stable growth state.
[0006] To achieve the above objectives, this utility model provides a multi-angle observation device with a multifunctional cell spheroidization culture dish, including a base, a shell cover, a battery pack, two light strips, a control board, multiple support devices, multiple reflectors, and a three-dimensional cell culture array;
[0007] The cover is mounted on the base, the battery pack is mounted inside the base, the two light strips are respectively mounted inside the base, the control board is mounted inside the base and connected to the two light strips and the control board, the multiple support devices are respectively mounted inside the base, the multiple reflectors are respectively mounted on the multiple support devices, and the three-dimensional cell culture array is mounted inside the base.
[0008] The three-dimensional cell culture array includes culture dish one, culture dish two, culture dish three, and culture dish four, all of which are installed inside the base.
[0009] Among them, culture dish one, culture dish two, culture dish three and culture dish four are all three-well.
[0010] Among them, culture dish one and culture dish three have four wells, and culture dish two and culture dish four have six wells.
[0011] The first culture dish has four wells, the second culture dish has six wells, and the third and fourth culture dishes each have three wells.
[0012] This utility model discloses a multi-angle observation device with a multifunctional cell spheroidization culture dish, comprising a base, a shell cover, a battery pack, two light strips, a control board, multiple support devices, multiple reflectors, and a three-dimensional cell culture array. The shell cover is mounted on the base, the battery pack is mounted inside the base, the two light strips are respectively mounted inside the base, the control board is mounted inside the base and connected to the two light strips and the control board, the multiple support devices are respectively mounted inside the base, the multiple reflectors are respectively mounted on the multiple support devices, and the three-dimensional cell culture array is mounted inside the base. The base provides installation conditions for the shell cover, battery pack, two light strips, fabrication plate, and multiple support devices. Multiple reflectors can be mounted via the support devices. The light source intensity can be adjusted via the battery pack, two light strips, and the control board. The reflectors reflect images of different sides of the three-dimensional cells. The images in the reflectors are then observed using a microscope for photographing. This allows for in-situ, multi-angle imaging of the sample without altering the microscope objective or sample orientation, without requiring external imaging devices, and while maintaining the three-dimensional cell growth state. The multifunctional cell culture dish and multi-angle observation device of this invention integrate three-dimensional cell culture with bottom and side observation functions, effectively avoiding the drawback of contamination caused by the reflector being too close to the cells, and supporting three-dimensional cell culture in hydrogels. During operation, cells are seeded onto the three-dimensional cell culture array and placed within the base for bottom and side microscopic imaging. The base also provides illumination for side imaging. Closing the shell cover during culture and imaging minimizes the risk of contamination. This approach achieves both lateral observation of three-dimensional cells and ensures cell growth is free from external interference. It significantly reduces sample contamination and the difficulty of imaging, allowing even untrained users to perform lateral imaging of three-dimensional cells. It enables in-situ, undisturbed lateral observation of three-dimensional cells without requiring expensive equipment, successfully overcoming the challenge of existing microscopy systems' inability to simultaneously cultivate in-situ samples during 3D sample observation. This provides an efficient, practical, and economical solution for cell research, powerfully promoting in-depth research and technological breakthroughs. It solves the problem that existing methods cannot simultaneously maintain a stable growth environment for in-situ observation of lateral morphological changes in three-dimensional cells. Attached Figure Description
[0013] To more clearly illustrate the technical solutions in the embodiments of this application or the prior art, the accompanying drawings used in the description of the embodiments or the prior art will be briefly introduced below.
[0014] Figure 1 This is a schematic diagram of the structure of a multi-angle observation device with a multifunctional cell spheroidization culture dish according to the first embodiment of this utility model.
[0015] Figure 2 This is a schematic diagram of the structure of a multi-angle observation device without a shell cover, which is based on the first embodiment of this utility model and includes a multifunctional cell spheroidization culture dish.
[0016] Figure 3 This is a schematic diagram of the battery pack structure shown in the first embodiment of this utility model.
[0017] Figure 4 This is a schematic diagram of the structure of a multi-angle observation device with a multifunctional cell spheroidization culture dish, according to the second embodiment of this utility model.
[0018] Figure 5 This is a schematic diagram of the structure of a multi-angle observation device without a shell cover, which is based on the second embodiment of this utility model and includes a multifunctional cell spheroidization culture dish.
[0019] Figure 6 This is a schematic diagram of the battery pack structure shown in the second embodiment of this utility model.
[0020] Figure 7 This is a schematic diagram of the structure of a multi-angle observation device with a multifunctional cell spheroidization culture dish according to the third embodiment of this utility model.
[0021] Figure 8 This is a schematic diagram of the structure of a multi-angle observation device without a shell cover, which is based on the third embodiment of this utility model and includes a multifunctional cell spheroidization culture dish.
[0022] Figure 9 This is a schematic diagram of the battery pack structure shown in the third embodiment of this utility model.
[0023] In the diagram: 101-base, 102-shell cover, 103-battery pack, 104-lamp strip, 105-control board, 106-support device, 107-reflector, 108-three-dimensional cell culture array, 109-culture dish one, 110-culture dish two, 111-culture dish three, 112-culture dish four. Detailed Implementation
[0024] The embodiments of this utility model are described in detail below. Examples of these embodiments are shown in the accompanying drawings, wherein the same or similar reference numerals denote the same or similar elements or elements having the same or similar functions throughout. The embodiments described below with reference to the accompanying drawings are exemplary and intended to explain this utility model, and should not be construed as limiting this utility model.
[0025] The first embodiment of this application is as follows:
[0026] Please see Figures 1-3This utility model provides a multi-angle observation device with a multifunctional cell spheroidization culture dish, including a base 101, a shell cover 102, a battery pack 103, two light strips 104, a control board 105, multiple support devices 106, multiple reflectors 107 and a three-dimensional cell culture array 108.
[0027] The cover 102 is mounted on the base 101, the battery pack 103 is mounted inside the base 101, the two light strips 104 are respectively mounted inside the base 101, the control board 105 is mounted inside the base 101 and connected to the two light strips 104 and the control board 105, the multiple support devices 106 are respectively mounted inside the base 101, the multiple reflectors 107 are respectively mounted on the multiple support devices 106, and the three-dimensional cell culture array 108 is mounted inside the base 101.
[0028] The three-dimensional cell culture array 108 includes culture dish 109, culture dish 210, culture dish 311 and culture dish 412, all of which are installed in the base 101.
[0029] Culture dish 109, culture dish 210, culture dish 311, and culture dish 412 are all three-well.
[0030] In this embodiment, the reflector 107 is fixed on the three-dimensional cell culture array 108. The three-dimensional cell culture array 108 and the reflector 107 can move together. The three-dimensional cell culture array can be used alone to photograph the three-dimensional cells in the array. The size of the three-dimensional cell culture array 108 with the reflector 107 can be adjusted according to different usage requirements. The material used to make it should be an ultra-low adhesion material with good biocompatibility. The three-dimensional cell culture array 108 can be placed in a housing with a light source for three-dimensional cell culture and observation. The housing includes two parts: a cover 102 and a base 101. The base 101 houses the light strip 104, battery, circuit board, and switch for adjusting the light intensity. Only the side of the base 101 closest to the three-dimensional cell culture array 108 is transparent and is made of a material with good light transmittance and high biocompatibility. The remaining parts are made of an opaque material with good biocompatibility. An inverted microscope is used in conjunction with the three-dimensional cell culture observation array in the first embodiment to perform three-dimensional cell culture and monitor three-dimensional cell growth.
[0031] Add 1×10 to each hole in the array 4A number of human prostate cancer cells (DU 145) were cultured in 100 μL of complete culture medium around the array to maintain cell growth. The three-dimensional cell culture observation array was placed in the base 101 and the cover 102 was placed on top. The entire array was placed on the microscope sample platform, the microscope's built-in light source was turned on, and the light source was adjusted to a suitable brightness. The sample state in the microscope eyepiece was observed, and the coarse and fine focus knobs of the microscope were adjusted sequentially until a clear image appeared in the field of view. Different magnification microscope objectives were used to take images of the bottom surface of the three-dimensional cells. The microscope objective was switched back to a larger field of view, and the sample stage was moved to find the side image of the sample in the reflecting mirror 107 in the eyepiece. The sample state in the microscope eyepiece was observed, and the coarse and fine focus knobs of the microscope were adjusted sequentially until a clear image of the side surface of the sample appeared in the field of view. Different magnification microscope objectives were used to take images of the bottom surface of the three-dimensional cells. The entire array was placed in a carbon dioxide incubator for culture, and the sample was taken out and photographed every 12 hours.
[0032] The second embodiment of this application is as follows:
[0033] Please see Figures 4-6 This utility model provides a multi-angle observation device with a multifunctional cell spheroidization culture dish, including a base 101, a shell cover 102, a battery pack 103, two light strips 104, a control board 105, multiple support devices 106, multiple reflectors 107 and a three-dimensional cell culture array 108.
[0034] The cover 102 is mounted on the base 101, the battery pack 103 is mounted inside the base 101, the two light strips 104 are respectively mounted inside the base 101, the control board 105 is mounted inside the base 101 and connected to the two light strips 104 and the control board 105, the multiple support devices 106 are respectively mounted inside the base 101, the multiple reflectors 107 are respectively mounted on the multiple support devices 106, and the three-dimensional cell culture array 108 is mounted inside the base 101.
[0035] The three-dimensional cell culture array 108 includes culture dish 109, culture dish 210, culture dish 311 and culture dish 412, all of which are installed in the base 101.
[0036] The first culture dish 109 and the third culture dish 111 have four wells, while the second culture dish 110 and the fourth culture dish 112 have six wells.
[0037] In this embodiment, an inverted microscope is used in conjunction with the three-dimensional cell culture observation array in the first embodiment to perform three-dimensional cell culture and monitor the growth of three-dimensional cells in the hydrogel.
[0038] Take 450 μL of melted matrix gel and 9 × 10⁻⁶ ppm in an ice box. 4 A number of human prostate cancer cells (DU 145) were mixed to a liquid volume of 990 μL, at which point the volume ratio of matrix gel to cell culture medium was 6:5; 110 μL of the mixture was added to each array well, at which point each well contained 1 × 10⁶ cells. 4 A number of human prostate cancer cells were observed. The three-dimensional cell culture observation array was placed in base 101, and the shell cover 102 was placed on top. The array was then incubated in a 37°C CO2 incubator for 30 minutes to allow the cell matrix gel to solidify. After 30 minutes, 50 μL of cell culture medium was added to each well, and the shell cover 102 was replaced. The entire array was placed on the microscope sample platform. The microscope's built-in light source was turned on and adjusted to a suitable brightness. The sample state in the microscope eyepiece was observed, and the coarse and fine focus knobs were adjusted sequentially until a clear image appeared in the field of view. Different magnification microscope objectives were used to photograph the bottom surface of the three-dimensional cells. The microscope objective was switched back to a larger field of view, and the sample stage was moved to find the side image of the sample in the reflecting mirror 107 in the eyepiece. The sample state in the microscope eyepiece was observed, and the coarse and fine focus knobs were adjusted sequentially until a clear image of the side surface of the sample appeared in the field of view. Different magnification microscope objectives were used to photograph the bottom surface of the three-dimensional cells. The entire array was placed in a CO2 incubator for culture, and photographed every 12 hours.
[0039] The third embodiment of this application is as follows:
[0040] Please see Figures 7-9 This utility model provides a multi-angle observation device with a multifunctional cell spheroidization culture dish, including a base 101, a shell cover 102, a battery pack 103, two light strips 104, a control board 105, multiple support devices 106, multiple reflectors 107 and a three-dimensional cell culture array 108.
[0041] The cover 102 is mounted on the base 101, the battery pack 103 is mounted inside the base 101, the two light strips 104 are respectively mounted inside the base 101, the control board 105 is mounted inside the base 101 and connected to the two light strips 104 and the control board 105, the multiple support devices 106 are respectively mounted inside the base 101, the multiple reflectors 107 are respectively mounted on the multiple support devices 106, and the three-dimensional cell culture array 108 is mounted inside the base 101.
[0042] The three-dimensional cell culture array 108 includes culture dish 109, culture dish 210, culture dish 311 and culture dish 412, all of which are installed in the base 101.
[0043] The first culture dish 109 has four wells, the second culture dish 110 has six wells, and the third culture dish 111 and the fourth culture dish 112 both have three wells.
[0044] In this embodiment, an inverted microscope is used in conjunction with the three-dimensional cell culture array 108 of the second embodiment, placed on the outer shell of the side observation base 101, to perform three-dimensional cell culture and obtain bright-field and DiO and DAPI stained fluorescence images of the bottom surface of the three-dimensional cell spheres. The stained cell spheres are transferred to the bottom of the three-dimensional cell culture array 108 in the dark, and 200 μL of buffer solution is added to achieve the best imaging effect. The inverted fluorescence microscope is turned on, and the built-in light source is adjusted to a suitable brightness. The image of the bottom surface of the three-dimensional cell sphere is found in a large field of view. Different magnification microscope objectives are used to obtain bright-field and fluorescence stained images of the bottom surface of the three-dimensional cell spheres. The microscope objective is adjusted to a large field of view, and the image of the three-dimensional cell sphere in the reflecting mirror 107 is found in the field of view. The built-in fluorescence light source of the microscope is adjusted to a suitable brightness, and the image of the bottom surface of the three-dimensional cell sphere is found in a large field of view. Different magnification microscope objectives are used to obtain fluorescence stained images of the sides of the three-dimensional cell spheres. The built-in light source of the microscope is turned off, the outer shell light source is turned on, and different magnification microscope objectives are used to obtain bright-field images of the sides of the three-dimensional cell spheres.
[0045] Beneficial effects:
[0046] I. It achieves undisturbed in-situ monitoring of the sides of three-dimensional cell spheres, with no direct contact between the device and the cell growth environment, resulting in minimal impact on the growth of three-dimensional cells (the mirror part is outside the cell culture array).
[0047] Second, the device achieves environmental stability during the imaging process (the shell and base are closed to capture three-dimensional cells without the cells coming into contact with the external environment).
[0048] Third, it fulfills the need for multi-angle observation while three-dimensional cells are cultured in a hydrogel scaffold (by seeding cells and hydrogel in a three-dimensional cell culture array).
[0049] The above-disclosed embodiments are merely one or more preferred embodiments of this application and should not be construed as limiting the scope of this application. Those skilled in the art can understand that all or part of the processes for implementing the above embodiments and equivalent changes made in accordance with the claims of this application still fall within the scope of this application.
Claims
1. A multi-angle observation device with a multifunctional cell spheroidization culture dish, characterized in that, It includes a base, a shell, a battery pack, two light strips, a control board, multiple support devices, multiple reflectors, and a three-dimensional cell culture array; The cover is mounted on the base, the battery pack is mounted inside the base, the two light strips are respectively mounted inside the base, the control board is mounted inside the base and connected to the two light strips and the control board, the multiple support devices are respectively mounted inside the base, the multiple reflectors are respectively mounted on the multiple support devices, and the three-dimensional cell culture array is mounted inside the base.
2. The multi-angle observation device with a multifunctional cell spheroidization culture dish as described in claim 1, characterized in that, The three-dimensional cell culture array includes culture dish one, culture dish two, culture dish three, and culture dish four, all of which are installed inside the base.
3. The multi-angle observation device with a multifunctional cell spheroidization culture dish as described in claim 2, characterized in that, Culture dish one, culture dish two, culture dish three, and culture dish four all have three wells.
4. The multi-angle observation device with a multifunctional cell spheroidization culture dish as described in claim 2, characterized in that, Culture dish one and culture dish three have four wells, while culture dish two and culture dish four have six wells.
5. The multi-angle observation device with a multifunctional cell spheroidization culture dish as described in claim 2, characterized in that, The first culture dish has four wells, the second culture dish has six wells, and the third and fourth culture dishes each have three wells.