Scanning ionic conductance microscope platform for single cell scanning and detection
By adding a protective device and a carbon dioxide supply device to the scanning ion conductivity microscope platform, the problems of unstable cell culture environment and optical interference were solved, and the stability and accuracy of cell scanning were achieved.
Patent Information
- Application Number
- CN202520214834.6
- Authority / Receiving Office
- CN · China
- Patent Type
- Utility models(China)
- Current Assignee / Owner
- Filing Date
- 2025-02-11
- Publication Date
- 2026-01-06
- Estimated Expiration
- 2035-02-11
AI Technical Summary
In existing scanning ion conductivity microscope platforms, the culture dishes are exposed to the air during cell imaging, which leads to an unstable cell culture environment, halo and shadow when illuminated by the light source, affecting the accuracy of cell positioning, and the probe tip is not clearly visible.
A protective device and a carbon dioxide supply device are added to the microscope platform. The protective device covers the culture dish to provide a closed environment. The gas environment is regulated by a carbon dioxide concentration detection device. The transparent cover and observation hole facilitate the probe to find target cells and reduce external interference.
A stable gaseous environment for in vitro cell culture was achieved, ensuring precise probe positioning of cells, reducing optical interference, and improving scanning accuracy and efficiency.
Smart Images
Figure CN223769998U_ABST
Abstract
Description
Technical Field
[0001] This invention belongs to the field of scanning ion conductivity microscopy technology, specifically relating to a scanning ion conductivity microscopy platform for single-cell scanning and detection. Background Technology
[0002] When cells are cultured in vitro, their survival environment includes nutrient supply, temperature, gaseous environment, liquid phase environment, and pH. The specific requirements for the cell survival environment are as follows:
[0003] Nutritional supply: Cells need basic nutrients such as amino acids, vitamins, and glucose, as well as cell growth factors, which are usually provided by serum.
[0004] Temperature: The culture temperature varies for tissues and cells from different biological sources. The most commonly used temperature for in vitro culture of animal cells is 37℃, while the standard temperature for human cell culture is 36.5℃±0.5℃.
[0005] Gaseous environment: A mixture of 5% CO2 and 95% air is required to provide oxygen. CO2 participates in the tricarboxylic acid cycle and maintains the pH of the culture medium.
[0006] Liquid environment: including water and balanced salt solutions, natural culture media (such as serum, tissue extracts) and synthetic culture media.
[0007] pH: The optimal pH range for most cells is 7.2 to 7.4. Deviations from this range will have a harmful effect on the cells.
[0008] Pollution-free and non-toxic: Maintain a non-toxic and sterile culture environment, promptly remove metabolites, and prevent cells from being contaminated or dying due to the accumulation of their own metabolic substances.
[0009] The cellular environment is crucial for a cell's growth, proliferation, and function. Suitable nutrient supply, temperature, gaseous environment, and pH are fundamental conditions for ensuring normal cell metabolism and growth. A clean and non-toxic environment ensures cell health and safety. These conditions work together to provide a stable and favorable environment for cell growth.
[0010] Current scanning ion conductivity microscopy platforms inevitably encounter problems during cell imaging:
[0011] The culture dish tray was exposed to the air, detached from the culture environment, which hindered continuous cell scanning and led to unsatisfactory experimental results. Furthermore,
[0012] The selected target cell is not necessarily located at the center of the culture dish. The culture dish tray needs to be moved to locate the suitable cell before scanning. However, during the process of light illuminating the objective lens, the light is affected by the probe platform, the bottom of the culture dish, the cells at the bottom of the dish, the liquid in the culture dish, and surrounding objects. This causes light reflection and refraction, resulting in halos and shadows around the cells observed through the eyepiece, and unclear imaging of the probe tip. These combined factors lead to a deviation in the relative position of the cell observed through the eyepiece and the probe tip. To avoid accidentally touching the cell and damaging the probe tip, the scanning range is expanded. Summary of the Invention
[0013] To address the problems existing in the prior art, the purpose of this invention is to provide a scanning ion conductivity microscope platform for single-cell scanning and detection. This invention can ensure the gaseous environment of cells during in vitro culture, while also facilitating the probe's search for target cells, based on existing scanning ion conductivity microscope platforms for single-cell scanning and detection.
[0014] To achieve the above objectives, the technical solution adopted by this utility model is as follows:
[0015] A scanning ion conductivity microscope platform for single-cell scanning and detection includes an inverted microscope body. A protective device is detachably connected to the culture dish tray of the inverted microscope body and covers the upper side of the culture dish. The protective device has an internal cavity for embedding the culture dish, which extends to the bottom of the protective device. A first observation hole is provided on the top of the protective device, which communicates with the top of the cavity. A transparent cover plate is provided on the upper side of the first observation hole, which can block the first observation hole and can move on the top of the protective device. A second observation hole is provided on the transparent cover plate for a probe mounted on the inverted microscope body to extend into. A carbon dioxide concentration detection device is provided above the culture dish in the cavity. The protective device also has a carbon dioxide input hole and a wire hole for the carbon dioxide concentration detection device. The protective device is connected to a carbon dioxide supply device through the carbon dioxide input hole.
[0016] Preferably, the first observation hole, the transparent cover plate, and the second observation hole are all circular in shape, the diameter of the transparent cover plate is larger than the diameter of the first observation hole, the diameter of the first observation hole is larger than the diameter of the second observation hole, and the diameter of the second observation hole is larger than the diameter of the probe.
[0017] Preferably, the protection device is arranged in a split type, including a first block and a second block with symmetrical structure, and the interface between the first block and the second block passes through the axis of symmetry of the protection device.
[0018] Preferably, both the first and second sections are detachably connected to the culture dish tray by screws, and both the first and second sections are provided with screw holes for the screws to pass through.
[0019] Preferably, the culture dish includes an inner cylinder and an outer cylinder, which are coaxially arranged. A water bath is formed between the outer side wall of the inner cylinder and the inner side wall of the outer cylinder. The water bath is equipped with a heating device and a temperature detection device. The cavity of the inner cylinder is used for culturing single cells. The outer side of the outer cylinder is provided with an outer edge plate. The outer diameter of the outer cylinder is not greater than the diameter of the embedding hole on the culture dish tray, and the outer diameter of the outer edge plate is greater than the diameter of the embedding hole.
[0020] Preferably, the inner and outer cylinders share the same petri dish bottom.
[0021] Preferably, the heating device and the temperature detection device are located on opposite sides of the water bath.
[0022] Preferably, a mounting ring is connected between the tops of the inner cylinder and the outer cylinder, and the mounting ring is used to install the heating device and the temperature detection device.
[0023] Preferably, the protection device has a heating device wire hole for the heating device wire to pass through, and a temperature detection device wire hole for the temperature detection device wire to pass through.
[0024] Preferably, the lower end of the probe holder of the inverted microscope body is provided with a light source, and the illumination direction of the light source is towards the objective lens of the inverted microscope body.
[0025] Compared with the prior art, the present invention has the following beneficial effects:
[0026] This invention relates to a scanning ion conductivity microscope platform for single-cell scanning and detection. Based on existing structures, it adds a protective device, a carbon dioxide supply device, and a carbon dioxide concentration detection device. The protective device, when installed on the culture dish tray, covers the culture dish, creating a relatively enclosed environment and reducing the impact of the external environment on the culture process. The protective device also provides insulation for the culture dish. The carbon dioxide supply device allows carbon dioxide to be introduced into the cavity of the protective device, and the carbon dioxide concentration detection device monitors the carbon dioxide concentration in the culture environment, providing a basis for adjusting the carbon dioxide supply. Therefore, this invention ensures a stable gaseous environment for cell culture in vitro. A first observation hole is located at the top of the protective device, providing space for the probe to locate target cells. When locating target cells, only the probe needs to be moved. The transparent cover seals the first observation hole, helping to maintain a stable culture environment. A second observation hole on the transparent cover facilitates probe insertion, ensuring normal probe detection. In summary, this invention can ensure the gaseous environment for cell culture in vitro, and also facilitates the probe's search for target cells. Attached Figure Description
[0027] To more clearly illustrate the technical solutions in the embodiments of this utility model, the drawings used in the description of the embodiments will be briefly introduced below. Obviously, the drawings described below are only some embodiments of this utility model. For those skilled in the art, other drawings can be obtained based on these drawings without creative effort.
[0028] Figure 1 This is a schematic diagram of the overall structure of an inverted microscope;
[0029] Figure 2 This is a first three-dimensional structural diagram of the protective device in an embodiment of this utility model;
[0030] Figure 3 This is a second three-dimensional structural diagram of the protective device in an embodiment of this utility model;
[0031] Figure 4 This is a schematic diagram of the first three-dimensional structure of the petri dish in an embodiment of this utility model;
[0032] Figure 5 This is a schematic diagram of the second three-dimensional structure of the petri dish in an embodiment of the present invention;
[0033] Figure 6 This is a schematic diagram of the probe holder structure in an embodiment of the present invention;
[0034] Figure 7 This is a first structural schematic diagram of the probe holder mounting position in an embodiment of this utility model;
[0035] Figure 8 This is a second structural schematic diagram of the probe holder mounting position in an embodiment of this utility model;
[0036] In the diagram, 1-light source, 2-objective lens, 3-eyepiece, 4-stage, 5-protective device, 5-1-first section, 5-2-second section, 5-3-screw hole, 5-4-first observation hole, 5-5-1-heating device wiring hole, 5-5-2-temperature detection device wiring hole, 5-6-1-carbon dioxide input port, 5-6-2-carbon dioxide concentration detection device wiring hole, 5-7-accommodating cavity, 6-transparent cover plate, 6 -1-Second observation hole, 7-Cultural dish, 7-1-Water bath, 7-2-Inner cylinder, 7-3-Outer cylinder, 7-4-Bottom of cultural dish, 7-5-Mounting ring, 7-6-Outer edge plate, 8-Probe holder, 9-Probe fixing knob, 10-Probe, 11-1-First lamp source, 11-2-Second lamp source, 12-Inverted microscope body, 13-Scanning ion conductivity microscope platform, 14-Cultural dish tray, 14-1-Embedding hole. Detailed Implementation
[0037] To make the objectives, technical solutions, and advantages of this utility model clearer, the embodiments of this utility model will be described in further detail below with reference to the accompanying drawings.
[0038] The accompanying drawings show various structural schematic diagrams according to embodiments of the present invention. These drawings are not to scale, and some details have been enlarged and may have been omitted for clarity. The shapes of the various regions and layers shown in the drawings, as well as their relative sizes and positional relationships, are merely exemplary and may deviate from reality due to manufacturing tolerances or technical limitations. Furthermore, those skilled in the art can design regions / layers with different shapes, sizes, and relative positions as needed.
[0039] Reference Figures 1-3 , Figure 7 and Figure 8 This embodiment uses a scanning ion conductivity microscope platform for single-cell scanning and detection. Based on an existing structure, it adds a protective device, a carbon dioxide supply device, and a carbon dioxide concentration detection device. Specifically, it includes an inverted microscope body 12. A protective device 5 is detachably connected to the culture dish tray 14 of the inverted microscope body 12, covering the upper side of the culture dish 7. The protective device 5 has an internal cavity 5-7 for embedding the culture dish 7. Figure 2Taking the orientation shown as an example, the cavity 5-7 extends to the bottom of the protective device 5. The top of the protective device 5 has a first observation hole 5-4, which communicates with the top of the cavity 5-7. When the culture dish 7 is placed in the embedding hole 14-1 on the culture dish tray 14, the protective device 5 is placed over the upper part of the culture dish 7, with the upper end of the culture dish 7 extending into the cavity 5-7. Then, the protective device 5 is connected to the culture dish tray 14, with the bottom of the protective device 5 tightly against the upper surface of the culture dish tray 14. The protective device 5 has a transparent cover 6 on the upper side of the first observation hole 5-4 (i.e., the top surface of the protective device 5) that can seal the first observation hole 5-4 and move on top of the protective device 5. The transparent cover 6 has a second observation hole 6-1 for the probe 10 mounted on the inverted microscope body 12 to extend into. The probe 10 can extend into the cavity 5-7 through the second observation hole 6-1 and approach the surface of the culture medium in the culture dish 7 to approach the target cells. In the above scheme, the transparent cover 6... The purpose of the transparent cover plate 6 sealing the first observation hole 5-4 is to stabilize the environment in the cavity 5-7 as much as possible, thereby ensuring the stability of the culture environment in the culture dish 7. The purpose of the movable transparent cover plate 6 is that when the probe 10 is used to find target cells, it can be found simply by moving the probe 10. When the probe 10 is searching for the target, the transparent cover plate 6 can move with the probe 10, and at the same time, the transparent cover plate 6 must still keep the first observation hole 5-4 sealed during the movement. A carbon dioxide concentration detection device is provided above the culture dish 7 in the cavity 5-7. The carbon dioxide concentration detection device can detect the carbon dioxide concentration in the culture environment, thereby providing a basis for the control of carbon dioxide flow rate. The protective device 5 is also provided with a carbon dioxide input hole 5-6-1 and a carbon dioxide concentration detection device through hole 5-6-2. The protective device 5 is connected to a carbon dioxide supply device through the carbon dioxide input hole 5-6-1, and the carbon dioxide supply device can introduce carbon dioxide into the cavity of the protective device.
[0040] In addition, the protective device 5 can be made of transparent material, so that it can be observed when the probe 10 moves, in order to find the target cell.
[0041] As an optional embodiment of this utility model, see Figure 2 and Figure 3In this embodiment, the first observation hole 5-4, the transparent cover plate 6, and the second observation hole 6-1 are all circular. The diameter of the transparent cover plate 6 is larger than the diameter of the first observation hole 5-4, the diameter of the first observation hole 5-4 is larger than the diameter of the second observation hole 6-1, and the diameter of the second observation hole 6-1 is larger than the diameter of the probe 10. Furthermore, it should be noted that the size of the transparent cover plate 6 must be within the movement range of the probe 10, and the transparent cover plate 6 must always completely block the first observation hole 5-4. The second observation hole 6-1 allows the probe 10 to pass through for scanning, but the culture dish tray may need to be moved to locate suitable cells.
[0042] As a preferred embodiment of this utility model, see Figure 2 and Figure 3 In this embodiment, the protective device 5 is designed in a split configuration, which facilitates the placement of the culture dish 7 and the installation of the probe 10, while also ensuring a certain degree of airtightness. Specifically, the protective device 5 in this embodiment includes a first segment 5-1 and a second segment 5-2 with symmetrical structures. The interface between the first segment 5-1 and the second segment 5-2 passes through the axis of symmetry of the protective device 5. More preferably, when splitting the protective device 5, the splitting is performed along the central axis of the first observation hole 5-4.
[0043] As a preferred embodiment of this utility model, see Figure 2 and Figure 3 Based on the above embodiments, in this embodiment, the first segment 5-1 and the second segment 5-2 are detachably connected to the culture dish tray 14 by screws. The first segment 5-1 and the second segment 5-2 are provided with screw holes 5-3 for screws to pass through. The screw holes 5-3 need to be arranged on the outer periphery of the outer edge of the culture dish 7.
[0044] As a preferred embodiment of this utility model, see Figure 4 and Figure 5This embodiment also improves upon existing culture dishes. Specifically, based on the existing commercially available culture dishes used for scanning cells, an inner cylinder 7-2 is added. A water bath 7-4 is formed between the inner cylinder 7-2 and the side wall of the original culture dish, making the culture dish 7 in this embodiment equivalent to a small water bath. Combined with a protective device, a constant temperature of 37 degrees Celsius and a 5% carbon dioxide concentration environment can be achieved, ensuring optimal cell condition. Specifically, the culture dish 7 in this embodiment includes an inner cylinder 7-2 and an outer cylinder 7-3, which are coaxially arranged. A water bath 7-1 is formed between the outer side wall of the inner cylinder 7-2 and the inner side wall of the outer cylinder 7-3. The water bath 7-1 is equipped with a heating device and a temperature detection device. The cavity of the inner cylinder 7-2 is used for culturing single cells. An outer edge plate 7-6 is provided on the outer side of the outer cylinder 7-3. The outer diameter of the outer cylinder 7-3 is not greater than the diameter of the embedding hole 14-1 on the culture dish tray 14. The outer diameter of the outer edge plate 7-6 is greater than the diameter of the embedding hole 14-1. Therefore, by using the outer edge plate 7-6 to rest on the upper side of the culture dish tray 14, the entire culture dish 7 can be stably placed on the culture dish tray 14. When using the petri dish 7 in this embodiment, water is added to the water bath 7-1, and the water in the water bath 7-1 is heated by a heating device. The water temperature in the water bath is detected by a temperature detection device, and the heating temperature of the heating device is controlled to maintain the water temperature in the water bath 7-1 within a set range to meet experimental requirements. Correspondingly, in this embodiment, the cross-sectional shape of the receiving cavity 5-7 inside the protective device 5 can be rectangular, circular, or polygonal. It is necessary to ensure that the receiving cavity 5-7 can limit the outer edge of the outer edge plate 7-6, so that the entire petri dish 7 can be stably placed on the petri dish tray 14.
[0045] As a preferred embodiment of this utility model, based on the above embodiments, in this embodiment, the inner cylinder 7-2 and the outer cylinder 7-3 share a petri dish bottom 7-4, which can reduce light reflection and refraction.
[0046] As a preferred embodiment of the present invention, based on the above embodiments, in this embodiment, the heating device and the temperature detection device are arranged on opposite sides of the water bath 7-1, so as to ensure that the water temperature in the water bath 7-1 can all reach the preset temperature range.
[0047] In a preferred embodiment of this utility model, based on the above embodiments, in this embodiment, an mounting ring 7-5 is connected between the tops of the inner cylinder 7-2 and the outer cylinder 7-3. The mounting ring 7-5 is used to install the heating device and the temperature detection device. The mounting ring 7-5 provides an installation position for the heating device and the temperature detection device, and also serves to stabilize the inner cylinder 7-2 and the outer cylinder 7-3.
[0048] In a preferred embodiment of this utility model, the protective device 5 has a heating device wire passage hole 5-5-1 for the heating device wire to pass through, and a temperature detection device wire passage hole 5-5-2 for the temperature detection device wire to pass through. The dimensions of the heating device wire passage hole 5-5-1 and the temperature detection device wire passage hole 5-5-2 are clearance-fitted or overfitted with the corresponding wires to minimize heat and gas loss from these holes.
[0049] As a preferred embodiment of this utility model, see Figures 6-8 Based on the above embodiments, in this embodiment, a light source is provided at the lower end of the probe holder 8 of the inverted microscope body 12, and the illumination direction of the light source is towards the objective lens 2 of the inverted microscope body 12. Installing a light source at the lower end of the probe holder ensures that the light source always shines directly into the objective lens, reducing reflection and refraction, ensuring that the eyepiece observes the true position of the probe and cells, reducing halos and shadows, and ultimately achieving precise positioning, narrowing the scanning range, and reducing scanning time.
[0050] As a preferred embodiment of the above solution, in this embodiment, at the lower end of the probe bracket 8, the light sources are provided on both sides of the probe 10, namely the first light source 11-1 and the second light source 11-2.
[0051] As a preferred embodiment of the above solution, in this embodiment, the light source can be an LED light source.
[0052] Finally, it should be noted that the above embodiments are only used to illustrate the technical solution of this utility model and not to limit it. Although the utility model has been described in detail with reference to the above embodiments, those skilled in the art should understand that modifications or equivalent substitutions can still be made to the specific implementation of this utility model. Any modifications or equivalent substitutions that do not depart from the spirit and scope of this utility model should be covered within the protection scope of the claims of this utility model.
Claims
1. A scanning ion conductance microscope platform for single cell scanning and detection, comprising: The protection device (5) is arranged on the inverted microscope body (12) in a split type, and includes a first split block (5-1) and a second split block (5-2) which are symmetrically structured. The first split block (5-1) and the second split block (5-2) are connected to the culture dish tray (14) through screws in a detachable manner, and each of the first split block (5-1) and the second split block (5-2) is provided with a screw hole (5-3) through which the screw passes.
2. The scanning ion-conductance microscope platform for single-cell scanning and detection of claim 1, wherein, The culture dish (7) includes an inner cylinder (7-2) and an outer cylinder (7-3). The inner cylinder (7-2) and the outer cylinder (7-3) are coaxially arranged, and a water bath tank (7-1) is formed between the outer side wall of the inner cylinder (7-2) and the inner side wall of the outer cylinder (7-3). The water bath tank (7-1) is provided with a heating device and a temperature detection device. The cavity of the inner cylinder (7-2) is used for culturing single cells. The outer cylinder (7-3) is provided with an outer edge plate (7-6) on the outer side surface. The outer diameter of the outer cylinder (7-3) is not greater than the diameter of the embedded hole (14-1) on the culture dish tray (14), and the outer diameter of the outer edge plate (7-6) is greater than the diameter of the embedded hole (14-1).
3. The scanning ion-conductance microscope platform for single-cell scanning and detection of claim 1, wherein, The inner cylinder (7-2) and the outer cylinder (7-3) share a culture dish bottom (7-4).
4. The scanning ion-conductance microscope platform for single-cell scanning and detection of claim 3, wherein, The heating device and the temperature detection device are arranged on opposite sides in the water bath tank (7-1).
5. The scanning ion-conductance microscope platform for single-cell scanning and detection of claim 1, wherein, The top portions of the inner cylinder (7-2) and the outer cylinder (7-3) are connected with a mounting ring (7-5), which is used for arranging the heating device and the temperature detection device.
6. The scanning ion-conductance microscope platform for single-cell scanning and detection of claim 5, wherein, 7. The scanning ion-conductance microscope platform for single-cell scanning and detection of claim 5, wherein, 8. The scanning ion-conductance microscope platform for single-cell scanning and detection of claim 5, wherein, 9. The scanning ion-conductance microscope platform for single-cell scanning and detection of claim 5, wherein, The protection device (5) is provided with a heating device wire through hole (5-5-1) for the heating device wire to pass through and a temperature detection device wire through hole (5-5-2) for the temperature detection device wire to pass through.
10. The scanning ion-conductance microscope platform for single-cell scanning and detection of claim 1, wherein, The lower end of the probe support (8) of the inverted microscope body (12) is provided with a lamp source, and the irradiation direction of the lamp source is towards the objective lens (2) of the inverted microscope body (12).