Stretchable cell culture dish capable of real-time microscope observation and marker detection

Abstract

The utility model relates to the technical field of cell culture consumables, in particular to a dynamic stretching cell culture dish capable of real-time microscope observation and marker detection, which comprises a culture dish main body, a bottom membrane and a sensor, the bottom film covers the bottom surface of the culture dish main body; the culture dish main body can be stretched along a set direction, and the upper surface of the culture dish main body is provided with a culture pond penetrating through the upper surface and the lower surface; among the side walls around the culture pond, the side wall parallel to the stretching direction is a stretching side wall, and the side wall perpendicular to the stretching direction is a fixed side wall; two groups of clamping plates are symmetrically arranged in the culture pond, each group of clamping plates is arranged close to the fixed side wall, and a set distance is formed between each group of clamping plates and the fixed side wall; and the sensors are arranged between each group of clamping plates and the fixed side wall. Stretching along a set direction to implement mechanical stimulation on the cells; the bottom membrane with limited thickness is matched with the inverted microscope, so that the change condition of the cells after mechanical stimulation can be observed; the sensor can detect cell secretion factors in real time.

Description

Technical Field

[0001] This utility model relates to the field of cell culture consumables technology, specifically to a dynamically stretchable cell culture dish that allows for real-time microscopic observation and biomarker detection. Background Technology

[0002] The information disclosed in this background section is intended only to enhance the understanding of the overall background of this utility model and is not necessarily to be construed as an admission or in any way implying that such information constitutes prior art known to those skilled in the art.

[0003] During cell culture, the mechanical microenvironment has a significant impact on the development and regeneration of organs and tissues, as well as on various levels of life activities such as cell proliferation, migration, differentiation, and apoptosis. However, current cell stretching culture dishes make it difficult to observe the dynamic response of cells to stretching in real time.

[0004] Meanwhile, inverted microscopes are commonly used for dynamic cell observation. To observe the cell response to stretching stimuli in real time, the objective lens needs to be focused on the cultured cell layer, i.e., the bottom surface of the culture dish. However, conventional cell culture dishes are relatively thick, especially for microscopes with autofocus functions. When the bottom film is thick, the microscope's autofocus adjustment speed cannot keep up with the changes in the thickness of the bottom film, resulting in the inverted microscope's objective lens being unable to focus or increasing aberrations.

[0005] Furthermore, stretching can stimulate the release of factors from cells. When using electrochemical sensors for measurement, the sensors must be fixed in place and kept away from the cell culture area to avoid affecting cell growth. Therefore, current cell culture dishes cannot simultaneously measure the release of factors from cell culture. Utility Model Content

[0006] To address the aforementioned issues, this invention provides a dynamically stretched cell culture dish that allows for real-time microscopic observation and biomarker detection.

[0007] To achieve the above technical objectives, the present invention adopts the following technical solution:

[0008] A stretchable cell culture dish for real-time microscopic observation and biomarker detection includes a culture dish body, a bottom membrane, and a sensor;

[0009] The bottom membrane covers the bottom surface of the culture dish body;

[0010] The culture dish body can be stretched along a set direction, and a culture pool is provided on the upper surface of the culture dish body, penetrating the upper and lower surfaces; among the side walls around the culture pool, the side walls parallel to the stretching direction are called stretching side walls, and the side walls perpendicular to the stretching direction are called fixed side walls.

[0011] Two sets of clamps are symmetrically arranged in the culture tank. Each set of clamps is located close to a fixed side wall, and there is a set distance between each set of clamps and the fixed side wall.

[0012] The sensor is positioned between each set of clamps and the fixed sidewall.

[0013] In one or more embodiments, the substrate is made of polydimethylsiloxane (PDMS).

[0014] In one or more embodiments, the thickness of the substrate film is 20–100 μm.

[0015] In one or more embodiments, the body of the culture dish is made of polydimethylsiloxane (PDMS).

[0016] In one or more embodiments, each set of clamps includes two symmetrically arranged limiting plates, which are parallel to the fixed sidewall and connected to the tension sidewall.

[0017] Preferably, the sum of the widths of the two limiting plates in each set of clamps is less than the width of the fixed sidewall, so that the two limiting plates are spaced at a set distance.

[0018] Preferably, the limiting plate is at a set distance from the bottom surface of the culture tank.

[0019] In one or more embodiments, the thickness of the fixed sidewall is greater than the thickness of the tensile sidewall.

[0020] In one or more embodiments, the set distance between each set of clamps and the fixed sidewall is slightly less than the thickness of the sensor.

[0021] In one or more embodiments, the bottom of the sensor is attached to the upper surface of the base film;

[0022] Preferably, a wire is connected to the top of the sensor.

[0023] In one or more embodiments, the upper surface of the fixed sidewall is symmetrically provided with positioning holes that penetrate the upper and lower surfaces.

[0024] In one or more embodiments, the area of ​​the bottom film is slightly larger than the area of ​​the bottom surface of the culture dish body.

[0025] The beneficial effects of this utility model are as follows:

[0026] (1) In this invention, stretching the sidewall and bottom membrane along the set stretching direction can achieve mechanical stimulation of the cells at the bottom of the culture pool; using a polydimethylsiloxane (PDMS) bottom membrane with a thickness of 20-100μm in conjunction with an inverted microscope, the morphological changes of the cells at the bottom of the culture pool after being mechanically stimulated can be directly observed; at the same time, the secretory factors of the cells can be detected in real time through the sensor between the clamp and the fixed sidewall.

[0027] (2) In this invention, both the main body of the culture dish and the bottom membrane are made of polydimethylsiloxane (PDMS), which has high light transmittance, elasticity and biocompatibility. The biocompatibility of the main body of the culture dish and the bottom membrane enables cells to grow, proliferate and differentiate in the membrane and culture pool; at the same time, the elasticity enables the stretching to apply mechanical stimulation to the cells; the high light transmittance and the set thickness of the bottom membrane can meet the requirements of the inverted microscope for observation of cells at the bottom of the culture pool (the upper surface of the bottom membrane).

[0028] (3) Traditional stretch culture dishes lack sensors or use external sensors, making real-time detection of cell-released factors difficult. External sensors are susceptible to mechanical interference, leading to signal drift or structural damage. In this invention, the sensor is positioned using a clamp, cleverly avoiding the impact of mechanical stress on the detection unit. Cell-secreted factors diffuse rapidly through the culture medium to the sensor interface, enabling real-time measurement. Simultaneously, the interference fit further restricts sensor placement, facilitating sensor fixation and replacement, and solving the problem of sensors being unable to operate for extended periods. Furthermore, two sensor placement locations are provided within the culture tank, enabling simultaneous detection of multiple cell-secreted factors. Attached Figure Description

[0029] The accompanying drawings, which form part of this utility model, are used to provide a further understanding of this utility model. The illustrative embodiments of this utility model and their descriptions are used to explain this utility model and do not constitute an improper limitation of this utility model.

[0030] Figure 1 This is a schematic diagram of a stretchable cell culture dish that allows for real-time microscopic observation and biomarker detection; the red double-headed arrows in the diagram indicate the stretching direction.

[0031] Figure 2 A cross-sectional view of a stretchable cell culture dish that allows for real-time microscopic observation and biomarker detection;

[0032] Among them, 1-bottom membrane, 2-culture dish body, 3-sensor, 4-fixed sidewall, 5-stretching sidewall, 6-culture pool, 7-limiting plate, 8-positioning hole and 9-sensor wire. Detailed Implementation

[0033] It should be noted that the following detailed description is exemplary and intended to provide further explanation of the present invention. Unless otherwise specified, all technical and scientific terms used in this invention have the same meaning as commonly understood by one of ordinary skill in the art to which this invention pertains.

[0034] It should be noted that the terminology used herein is for the purpose of describing particular embodiments only and is not intended to limit the exemplary embodiments according to this utility model. As used herein, the singular form is intended to include the plural form as well, unless the context clearly indicates otherwise. Furthermore, it should be understood that when the terms "comprising" and / or "including" are used in this specification, they indicate the presence of features, steps, operations, devices, components, and / or combinations thereof.

[0035] The present invention will be further described in detail below with reference to specific embodiments. It should be noted that the specific embodiments are explanations of the present invention and not limitations.

[0036] Example 1

[0037] See Figure 1 and Figure 2 A stretchable cell culture dish for real-time microscopic observation and marker detection includes a culture dish body 2, a bottom membrane 1, and a sensor 3.

[0038] The bottom film 1 covers the bottom surface of the culture dish body 2;

[0039] The culture dish body 2 can be stretched on both sides along a set direction. A culture pool 6 is provided on the upper surface of the culture dish body 2, penetrating the upper and lower surfaces. The two side walls parallel to the stretching direction of the culture pool 6 are called stretching side walls 5, and the side walls perpendicular to the stretching direction are called fixed side walls 4.

[0040] Two sets of clamps are symmetrically arranged inside the culture tank 6. Each set of clamps is located close to the fixed side wall 4, and the distance between each set of clamps and the fixed side wall 4 is set.

[0041] Sensor 3 is positioned between each set of clamps and the fixed side wall 4.

[0042] The bottom membrane 1 is made of polydimethylsiloxane (PDMS), with a thickness of 20–100 μm. The thickness of the bottom membrane 1 is set according to the actual needs of observation under an inverted microscope and the stress limit. The culture dish body 2 is also made of PDMS, and the proportion of PDMS curing agent used is adjusted according to the stretching requirements. Both the culture dish body 2 and the bottom membrane 1 are made of PDMS, which has high light transmittance, elasticity, and biocompatibility. The biocompatibility of the culture dish body and the bottom membrane allows cells to grow, proliferate, and differentiate in the membrane and culture tank; its elasticity allows for stretching to apply mechanical stimulation to the cells; and the high light transmittance and set thickness of the bottom membrane meet the requirements for observation of cells at the bottom of the culture tank (on the upper surface of the bottom membrane) under an inverted microscope.

[0043] Each set of clamps includes two symmetrically arranged limiting plates 7, which are parallel to the fixed sidewall 4 and connected to the tension sidewall 5. The limiting plates 7 are integrally formed with the culture dish body 2, and the material of the limiting plates 7 is the same as that of the culture dish body 2, which is polydimethylsiloxane (PDMS).

[0044] The sum of the widths of the two limiting plates 7 in each set of clamps is less than the width of the fixed sidewall 4, so that there is a set distance between the two limiting plates 7. The set distance between the two limiting plates 7 can not only satisfy the limiting of the sensor, but also increase the contact area between the sensor and the culture medium, thereby increasing the accuracy of detecting factors secreted by cells in the culture medium.

[0045] The limiting plate 7 is at a set distance from the bottom surface of the culture tank 6. The limiting plate 7 does not directly contact the bottom surface of the culture tank 6 and will not affect the growth of cells cultured on the bottom surface of the culture tank 6, i.e., the upper surface of the bottom membrane 1.

[0046] When not stretched, the set distance between each set of clamps and the fixed sidewall 4 is slightly less than the thickness of the sensor 3. The sensor 3 and each set of clamps and the fixed sidewall 4 are interference-fitted, so that the sensor 3 can be firmly confined between each set of clamps 7 and the fixed sidewall 4. When placing or removing the sensor 3, only the clamps 7 need to be stretched. During the actual stretching process, when the stretchable cell culture dish, which allows for real-time microscopic observation and marker detection, is stretched along the set direction with the cell analyzer, only the stretching sidewall 5 and the bottom membrane 1 undergo slight deformation (deformation range 0.1%-25%, such as 0.25mm-10mm when the total length of the culture dish is 40mm). The deformation of each set of clamps that confines the sensor is very small and can be ignored. Therefore, during the stretching process, the sensor 3 will not change position or move due to excessive deformation of the clamps 7.

[0047] Traditional stretched culture dishes lack sensors or use external sensors, making real-time detection of cell-released factors difficult. External sensors are susceptible to mechanical interference, leading to signal drift or structural damage. In this embodiment, a clamp is used to limit the sensor's position, cleverly avoiding the impact of mechanical stress on the detection unit. Cell-secreted factors rapidly diffuse through the culture medium to the sensor interface, enabling real-time measurement. Simultaneously, interference fits are used to limit the sensor's position, facilitating sensor fixation and replacement, and solving the problem of sensors being unable to operate for extended periods. Furthermore, two sensor placement locations are provided within the culture tank, enabling simultaneous detection of multiple cell-secreted factors.

[0048] To ensure better support and stability of the sidewalls during the stretching process and reduce the risk of deformation and breakage, and to ensure smooth stretching, the thickness of the fixed sidewall 4 is greater than the thickness of the stretching sidewall 5.

[0049] The bottom of sensor 3 is attached to the upper surface of the bottom membrane 1; sensor 3 can accurately detect the factors secreted by the cells cultured in culture tank 6.

[0050] For signal transmission / power supply, a wire 9 is connected to the top of the sensor 3.

[0051] In order to enable real-time microscopic observation and marker detection, when the stretchable cell culture dish is stretched along a set direction in conjunction with the cell instrument, the upper surface of the fixed sidewall is symmetrically provided with positioning holes 8 that penetrate the upper and lower surfaces. The positioning holes 8 can cooperate with the stretching of the cell instrument to realize the stretching of the culture dish.

[0052] To illustrate in detail the relationship between the positioning hole and the cell strainer stretching device in this application, we will take the high-precision cell culture stretching device for dynamic microscopic observation (hereinafter referred to as the stretching device) disclosed in Chinese Patent CN219950959 U as an example.

[0053] The "stretchable cell culture dish capable of real-time microscopic observation and biomarker detection" of this utility model is placed in the cell culture pool of the stretching apparatus. By passing the positioning pin through the positioning hole on the stretching support and the fixing support in the stretching apparatus, as well as the positioning hole 8 in the "stretchable cell culture dish capable of real-time microscopic observation and biomarker detection" of this utility model, the "stretchable cell culture dish capable of real-time microscopic observation and biomarker detection" of this utility model can be fixed on the stretching apparatus disclosed in Chinese Patent CN 219950959 U, thereby enabling subsequent stretching culture.

[0054] To ensure a tight bond between the bottom membrane 1 and the culture dish body 2, the area of ​​the bottom membrane 1 is slightly larger than the area of ​​the bottom surface of the culture dish body 2 during preparation.

[0055] Finally, it should be noted that the above description is merely a preferred embodiment of this utility model and is not intended to limit this utility model. Although this utility model has been described in detail with reference to the foregoing embodiments, those skilled in the art can still modify the technical solutions described in the foregoing embodiments or make equivalent substitutions for some of them. Any modifications, equivalent substitutions, improvements, etc., made within the spirit and principles of this utility model should be included within the protection scope of this utility model.

Claims

1. A stretchable cell culture dish that enables real-time microscopic observation and marker detection, characterized by, This includes the petri dish body, the bottom membrane, and the sensor; The bottom membrane covers the bottom surface of the culture dish body; The culture dish body can be stretched along a set direction, and a culture pool is provided on the upper surface of the culture dish body, penetrating the upper and lower surfaces; Among the side walls around the culture tank, the side walls parallel to the stretching direction are called stretching side walls, and the side walls perpendicular to the stretching direction are called fixed side walls. Two sets of clamps are symmetrically arranged in the culture tank. Each set of clamps is located close to a fixed side wall, and there is a set distance between each set of clamps and the fixed side wall. The sensor is positioned between each set of clamps and the fixed sidewall.

2. The stretchable cell culture dish of claim 1, wherein, The substrate is made of polydimethylsiloxane; the thickness of the substrate is 20–100 μm.

3. The stretchable cell culture dish of claim 1, wherein, The main body of the culture dish is made of polydimethylsiloxane.

4. The stretchable cell culture dish of claim 1, wherein, Each set of clamps includes two symmetrically arranged limiting plates, which are parallel to the fixed sidewall and connected to the tension sidewall.

5. The stretchable cell culture dish of claim 4, wherein, The sum of the widths of the two limiting plates in each set of clamps is less than the width of the fixed sidewall, so that the two limiting plates are spaced at a set distance. The limiting plate is at a set distance from the bottom surface of the culture tank.

6. The stretchable cell culture dish of claim 1, wherein, The thickness of the fixed sidewall is greater than the thickness of the tensile sidewall.

7. The stretchable cell culture dish of claim 1, wherein, The set distance between each set of clamps and the fixed sidewall is slightly less than the thickness of the sensor.

8. The stretchable cell culture dish as described in claim 1, characterized in that, The bottom of the sensor is attached to the upper surface of the bottom membrane; The sensor has a wire connected to its top.

9. The stretchable cell culture dish of claim 1, wherein, The upper surface of the fixed sidewall is symmetrically provided with positioning holes that penetrate the upper and lower surfaces.

10. The stretchable cell culture dish of claim 1, wherein, The area of ​​the bottom membrane is slightly larger than the area of ​​the bottom surface of the culture dish.

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