Cell culture device

Abstract

The utility model belongs to the field of life science, and relates to a cell culture device which comprises a supporting frame and a cell culture plate laid on the supporting frame, and imaging holes are formed in the cell culture plate. The utility model provides a cell culture device which is simple in structure, convenient to assemble and easy to operate.

Description

Technical Field

[0001] This utility model belongs to the field of life sciences and relates to a cell culture device, and more particularly to a cell culture device adapted to a lensless imaging mode. Background Technology

[0002] Lensless microscopy is a novel microscopic method distinct from traditional lenticular microscopes. In traditional lenticular microscopes, lenses are the core of the imaging process, focusing and magnifying the object to obtain its microscopic structure. The complex optical path design and lens groups make miniaturization difficult, and high magnification leads to a drastic reduction in the field of view. In lensless microscopy, the light source, sample, and sensor are aligned on a straight optical path. The microscope directly records the projection of the object onto the photoelectric sensor under the light source, and then uses appropriate algorithms to generate a full-field image. Because this emerging imaging mode eliminates optical components such as lens groups, the overall microscope structure is simple and compact, and it is not constrained by the contradictory relationship between magnification and field of view, allowing for a wider field of view while maintaining high object resolution.

[0003] These characteristics make lensless microscopy widely applicable in life sciences, especially in large-scale cell analysis in in situ environments. Its miniaturized size makes it easier to place in cell culture chambers, enabling continuous in situ monitoring of cells and providing rich information on the dynamic processes of cell growth, migration, differentiation, and apoptosis (Zheng Guoan, et al. PNAS, 2011, 108(41): 16889-16894). Furthermore, at the same resolution, lensless imaging instruments achieve a field of view far exceeding that of lensed microscopes, allowing for high-throughput cell detection and screening. For example, patent application CN112131945A discloses a deep learning-based lensless microscopy technique, combined with microfluidic technology, to achieve high-throughput, large-volume statistical observation of samples.

[0004] With the development of cell biology, it has been discovered that extensive interactions exist between different cell types in the body, playing a crucial role in cell proliferation, differentiation, and apoptosis. Co-culturing multiple cell types can better simulate the in vivo microenvironment and provide a deeper understanding of life science processes. Based on different culture methods, cell co-culture can be divided into direct contact co-culture and indirect contact co-culture. In indirect contact co-culture, the two types of cells do not directly contact each other; instead, they communicate and interact across cells through the diffusion of signaling molecules. This method results in low chaos and controllable variables throughout the process, making it the most widely used co-culture system currently.

[0005] Currently, mainstream indirect contact co-culture devices, such as the Transwell chamber culture system, generally consist of upper and lower chambers. The bottom of the upper chamber has a molecularly permeable polycarbonate membrane that connects the upper and lower chambers. The two types of cells seeded in the upper and lower chambers do not directly contact each other; instead, signal molecule transmission occurs through the semi-permeable membrane. However, in lensless imaging methods, the Transwell chamber membrane and its supporting structure can obstruct the imaging light source, making it difficult to obtain good images. To date, a multi-cell co-culture device suitable for lensless microscopy has not been developed. The use of lensless imaging systems remains limited to single-species cell research. This significantly restricts the application of lensless imaging to the study of deeper and more complex cellular activities.

[0006] In conclusion, existing lensless imaging devices cannot meet the requirements of deeper cell interaction research, and there is an objective need to design and manufacture a cell culture device that is compatible with it. Utility Model Content

[0007] In order to solve the above-mentioned technical problems in the background art, the present invention provides a cell culture device that is simple in structure, easy to assemble and easy to operate.

[0008] To achieve the above objectives, the present invention adopts the following technical solution: a cell culture device, the cell culture device including a support frame and a cell culture plate laid on the support frame, the cell culture plate being provided with an imaging hole.

[0009] Preferably, the imaging hole used in this invention penetrates the cell culture plate along the thickness direction of the cell culture plate.

[0010] Preferably, the imaging aperture used in this invention is obtained by extending radially outward from the geometric center of the cell culture plate.

[0011] Preferably, the imaging aperture used in this invention is a rectangular aperture; the four corners of the rectangular aperture are provided with rounded chamfers.

[0012] Preferably, the support frame used in this invention has a hollow cylindrical structure, and the cell culture plate is laid on the support frame radially.

[0013] Preferably, the inner wall of the support frame used in this utility model is provided with a boss, the outer diameter of the cell culture plate is smaller than the inner diameter of the support frame, and the cell culture plate is arranged on the boss along the radial direction of the support frame.

[0014] Preferably, the boss used in this utility model is in the form of a ring or a block; when the boss is in the form of a block, there are multiple bosses, the upper surfaces of the multiple bosses are on the same plane, and the multiple bosses are evenly distributed on the inner wall of the support frame along the radial direction of the support frame.

[0015] Preferably, the support frame used in this utility model is provided with a working window that penetrates the side wall of the support frame; the working window extends downward from the top of the support frame along the axial direction of the support frame, and there are one or more working windows.

[0016] Preferably, the support frame used in this utility model is provided with support feet at the bottom, and there are three support feet, which are evenly distributed at the bottom of the support frame.

[0017] Preferably, the cell culture device provided by this utility model further includes a photoelectric sensor and a culture dish; the photoelectric sensor is placed at the bottom of the support frame; the photoelectric sensor is located at the vertical projection of the imaging hole; and the culture dish is covered outside the support frame.

[0018] The advantages of this utility model are:

[0019] This invention provides a cell culture device, including a support frame and a cell culture plate laid on the support frame, with an imaging hole provided on the cell culture plate. The inner wall of the support frame may have protrusions for supporting the cell culture plate, and support feet are provided at the bottom of the support frame. The cell culture plate is made of a highly transparent biocompatible material, and its shape conforms to the interior of the support frame. A hollow imaging hole is provided in the central area of ​​the cell culture plate, and the specific position and shape of the imaging hole are determined or influenced by the lensless imaging mode sensor. The cell culture plate has a central through-path design between the light source and the sensor, ensuring that the microscope's optical path is not affected by the cell culture plate, thus guaranteeing optimal image quality for the lensless microscope. This invention greatly expands the application of lensless imaging in cell research without compromising image quality. When using this cell culture device, the cell culture plate can be used as a general cell slide for cell seeding, cell passage, and other operations. Furthermore, cell culture plates can be removed or loaded at any time as needed. The high light transmittance of the cell culture plates ensures that further analysis of the cells on them is possible, including but not limited to: cell morphology and adhesion analysis, cell immunoassay staining, electrophysiological measurements, drug sensitivity tests, transcriptomics and proteomics measurements, demonstrating great flexibility and versatility. This invention provides a cell culture device that is compatible with lensless imaging mode. It has a simple structure, is easy to operate, and has significant economic advantages. It can expand applications related to multi-cell co-culture and cell-cell interaction research in lensless imaging mode. Attached Figure Description

[0020] Figure 1This is a schematic diagram of the overall structure of the cell culture device provided by this utility model;

[0021] Figure 2 This is a schematic diagram of the cell culture plate used in this utility model;

[0022] Figure 3 This is a schematic diagram of the support frame used in this utility model;

[0023] Figure 4 This is a schematic diagram of the cell culture device provided by this utility model installed in a lensless microscopy imaging system;

[0024] in:

[0025] 1-Cell culture plate; 2-Support frame; 3-Imaging hole; 4-Boss; 5-Working window; 6-Supporting foot; 7-Photoelectric sensor; 8-Cultural dish. Detailed Implementation

[0026] See Figure 1 This invention provides a cell culture device, including a support frame 2 and a cell culture plate 1 laid on the support frame 2, wherein the cell culture plate 1 is provided with an imaging hole 3. For example, the cell culture plate 1 used in this invention is made of highly transparent polystyrene or other biocompatible materials with high light transmittance. The cell culture device provided by this invention is suitable for lensless imaging modes of cell proliferation culture, cell scratch assay, cell invasion assay, cell co-culture, primary cell culture, and cell imaging staining.

[0027] See Figure 1 as well as Figure 2 The imaging hole 3 penetrates the cell culture plate 1 along its thickness direction. The imaging hole 3 extends radially outward from the geometric center of the cell culture plate 1. For example, the imaging hole 3 is a rectangular hole. Preferably, the rectangular hole used in this invention has rounded chamfers at its four corners. It should be noted that the specific position and shape of the imaging hole 3 are determined or influenced by the lensless sensor chip, and will not be elaborated further here.

[0028] See Figure 1 as well as Figure 3The support frame 2 used in this invention has a hollow cylindrical structure, and the cell culture plate 1 is laid on the support frame 2 radially. To stably lay the cell culture plate 1 on the support frame 2, this invention uses an embedded method, placing the cell culture plate 1 inside the support frame. For example, the inner wall of the support frame 2 is provided with protrusions 4, the outer diameter of the cell culture plate 1 is smaller than the inner diameter of the support frame 2, and the cell culture plate 1 is arranged radially on the protrusions 4. Preferably, the protrusions 4 are generally annular or block-shaped; when the protrusions 4 are block-shaped, there are multiple protrusions 4, the upper surfaces of the multiple protrusions 4 are on the same plane, and the multiple protrusions 4 are evenly distributed radially on the inner wall of the support frame 2. Regardless of the method, this provides support for the cell culture plate, facilitating its placement. When moving or transporting the support frame 2, because the cell culture plate 1 is embedded within the support frame 2, it will not tilt or slip, providing excellent stability. It should be noted that the distance from the boss 4 to the top of the support frame 2 is not less than the thickness of the cell culture plate 1.

[0029] To facilitate manipulation of the cell culture plate 1, such as removing it with tweezers, the support frame 2 used in this invention is provided with a working window 5 penetrating the side wall of the support frame 2. The working window extends downward from the top of the support frame 2 along the axial direction of the support frame 2. There are one or more working windows 5. When there are multiple working windows 5, the cell culture plate 1 can be operated from any direction, making it more convenient to use. For example, the length of the working window 5 can be 3-5 mm.

[0030] The bottom of the support frame 2 is provided with support feet 6. There are three support feet 6, which are evenly distributed at the bottom of the support frame 2.

[0031] See Figure 4 For example, the cell culture apparatus also includes a photoelectric sensor 7 and a culture dish 8; the photoelectric sensor 7 is placed at the bottom of the support frame 2; the photoelectric sensor 7 is located at the vertical projection of the imaging hole 3; the culture dish 8 is covered outside the support frame 2.

[0032] The cell culture apparatus provided by this utility model will be described exemplarily below with reference to the accompanying drawings:

[0033] Example 1

[0034] See Figure 1This invention provides a cell culture device, including a circular cell culture plate 1 and a circular support frame 2. The cell culture plate 1 is made of a transparent material, such as polystyrene. A rectangular imaging hole 3 is provided in the central area of ​​the substrate, and the four corners of the rectangle are rounded. A boss 4 is provided on the inner wall of the support frame 2, and the height of the boss 4 is equal to the thickness of the cell culture plate 1. A working window 5 is provided on the support frame 2 for easy installation and removal of the cell culture plate 1, and the working window 5 extends through the entire side wall of the support frame 2. Three equidistant support feet 6 are provided at the bottom of the support frame 2. See also Figure 4 The circular support frame 2 is positioned above the photoelectric sensor 7, with the imaging hole 3 on the cell culture plate 1 positioned directly above the photoelectric sensor 7 without any obstruction. For example, in this embodiment, the cell culture plate 1 has a diameter of 30 mm and a thickness of 1.5-2 mm. The geometric center of the rectangular imaging hole 3 in the center of the cell culture plate 1 coincides with the center of the circle of the cell culture plate 1, and the rectangle has a length and width of 12 mm.

[0035] Cells were seeded separately on the lensless imaging chip and the cell culture plate. The cell culture plate 1 could be placed separately in a culture dish for cell seeding, or it could be combined with the support frame 2 and seeded in a culture dish 8, ensuring that cells from the upper cell culture plate 1 did not seed the lower chip area. The high light transmittance of the cell culture plate 1 ensured that cell growth and development could be monitored using a standard bright-field microscope, and allowed for biochemical assays such as cell morphology and adhesion analysis, cell immunoassay staining, and electrophysiological measurements on the upper cells. See also... Figure 4 When using the device, add cell culture reagent to the culture dish 8, ensuring the reagent level is 5mm above the plane of the cell culture plate 1. The rectangular imaging hole 3 in the central area of ​​the cell culture plate 1 is vertically aligned with the photoelectric sensor 7, ensuring vertical transparency of the light source and preventing the culture plate from obstructing the lensless imaging light path and damaging the imaging quality.

[0036] The support frame 2 is equipped with a working window 5, into which clamps such as tweezers, clips or mini ophthalmic forceps can be inserted to facilitate the placement and handling of cell culture plates during experiments. This reduces damage to cells caused by difficulty in handling the culture plates, simplifies the separation of upper and lower cells, and allows for separate sampling of upper or lower cells, avoiding experimental result deviations due to cell contamination.

[0037] Cell culture plate 1 and support frame 2 can be recovered after the experiment and cleaned by soaking in trypsin solution, alcohol, isopropanol or other cleaning solutions, or by ultrasonic oscillation cleaning. The cleaned device can be reused.

[0038] Example 2

[0039] In this embodiment, the cell culture plate 1 can be designed with any thickness less than the height from the protrusion 4 to the top of the support frame 2 to match the needs of different biochemical experiments, such as 0.13-0.17 mm or 0.17–0.21 mm. Taking 0.13-0.17 mm as an example, it is widely used in most microscopic imaging experiments, suitable for high-power objectives (×60, ×100) working distances, and can be used for experiments such as immunofluorescence, calcium imaging, live cell imaging, and electrophysiological experiments. Taking 0.17-0.21 mm as an example, it can be used for cell mechanics-related experiments, such as cell stretching and shear stress-related experiments.

[0040] Example 3

[0041] In this embodiment, since the cell culture plate 1 is designed to be replaceable, multiple culture plates can be used to culture multiple types of cells at the same time. During the co-culture process, the types of cells in the upper layer can be changed according to the experimental arrangement and needs to construct specific inflammatory damage and disease models, such as immune cell repair models, immune response models or multi-cell dynamic turnover models in tissue repair.

Claims

1. A cell culture device, characterized in that: The cell culture device includes a support frame (2) and a cell culture plate (1) laid on the support frame (2). The cell culture plate (1) is provided with an imaging hole (3). The imaging hole (3) penetrates the cell culture plate (1) along the thickness direction of the cell culture plate (1). The imaging hole (3) is obtained by extending outward from the geometric center of the cell culture plate (1) along the radial direction of the cell culture plate (1). The imaging hole (3) is a rectangular hole. The four corners of the rectangular hole are provided with rounded chamfers.

2. The cell culture apparatus according to claim 1, characterized in that: The support frame (2) is a hollow cylindrical structure, and the cell culture plate (1) is laid on the support frame (2) radially.

3. The cell culture apparatus according to claim 2, characterized in that: The inner wall of the support frame (2) is provided with a boss (4), the outer diameter of the cell culture plate (1) is smaller than the inner diameter of the support frame (2), and the cell culture plate (1) is arranged on the boss (4) along the radial direction of the support frame (2).

4. The cell culture apparatus according to claim 3, characterized in that: The boss (4) is generally ring-shaped or block-shaped; when the boss (4) is block-shaped, there are multiple bosses (4), the upper surfaces of the multiple bosses (4) are on the same plane, and the multiple bosses (4) are evenly distributed on the inner wall of the support frame along the radial direction of the support frame (2).

5. The cell culture apparatus according to claim 4, characterized in that: The support frame (2) is provided with a working window (5) that penetrates the side wall of the support frame (2); the working window extends downward from the top of the support frame (2) along the axial direction of the support frame (2), and there are one or more working windows (5).

6. The cell culture apparatus according to claim 5, characterized in that: The bottom of the support frame (2) is provided with support feet (6), and there are four support feet (6) evenly distributed at the bottom of the support frame (2).

7. The cell culture apparatus according to claim 6, characterized in that: The cell culture device also includes a photoelectric sensor (7) and a culture dish (8); the photoelectric sensor (7) is placed at the bottom of the support frame (2); the photoelectric sensor (7) is located at the vertical projection of the imaging hole (3); the culture dish (8) is covered outside the support frame (2).

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