Cell climbing slide immunofluorescence staining device and method
By setting conical crawler grooves of different volumes on the staining plate of the cell crawler immunofluorescence staining device and using a film of hydrophobic material, the problem of uneven diffusion of antibodies is solved, the staining effect and test efficiency are improved, and the cost is reduced.
Patent Information
- Application Number
- CN202411711406.0
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2024-11-27
- Publication Date
- 2025-03-07
AI Technical Summary
In traditional cell climber immunofluorescence staining technology, there is material difference between the material of the cell culture plate used and the glass slide, resulting in uneven diffusion of antibodies, affecting the staining effect and increasing the cost of the test.
An immunofluorescence staining device for cell crawlers was designed, using four sets of conical crawler grooves of different volumes on the staining plate, and a film of hydrophobic material was set on the inner surface of the groove to evenly distribute specific fluorescent antibodies.
By evenly distributing antibodies, the staining effect is improved, the antibody usage is reduced, the test cost is reduced, and the specificity and contrast of staining is improved.
Smart Images
Figure CN119574267A_ABST
Abstract
Description
Technical Field
[0001] The invention belongs to the field of cell biology, and in particular relates to a cell slide immunofluorescence staining device and method. Background Art
[0002] Immunofluorescence staining is a technique that uses a combination of specific antibodies and fluorescent dyes to mark target proteins in cells or tissues. Through fluorescence microscopy, the target protein can be clearly located and quantitatively expressed;
[0003] The slide technology is to culture cells on a glass slide with a specific coating to form a monolayer of cells. The cell slide can better react with antibodies, improve the effect of fluorescent staining, and facilitate subsequent observation and analysis.
[0004] However, traditionally, cell culture plates or glass slides are usually used directly for cell staining. Cell culture plates are made of plastic and have a large difference in bottom area from the cell slide, which results in a large amount of fluorescent antibodies being used in the staining process. At the same time, glass slides are made of glass, which will produce infiltration during the staining process, causing uneven diffusion of antibodies, which in turn leads to uneven distribution of antibody concentration in the staining area, weakened staining effect, and increased experimental costs. Summary of the invention
[0005] The purpose of the present invention is to provide a cell slide immunofluorescence staining device and method to solve the problems raised in the above background technology.
[0006] To achieve the above object, the present invention provides the following technical solutions:
[0007] A cell slide immunofluorescence staining device comprises a main body, a staining plate, four groups of slide grooves arranged on the staining plate, and a hydrophobic material film arranged in the slide grooves; a connecting mechanism is arranged on the staining plate.
[0008] Preferably, the volumes of the four groups of climbing slice grooves are different, namely 10 microliters, 20 microliters, 40 microliters and 80 microliters, and the diameters of the bottoms of the four groups of climbing slice grooves are 10 mm, 16 mm, 22 mm and 26 mm, respectively.
[0009] Preferably, the hydrophobic material film is a PTFE film.
[0010] Preferably, the climbing sheet groove is configured with a conical round bottom, with a large upper opening and a small lower round bottom.
[0011] Preferably, the hydrophobic material film is located on the round bottom of the sheet climbing groove.
[0012] Preferably, the hydrophobic material film is located on the conical inner wall of the sheet climbing groove.
[0013] Preferably, a concentration indicator bar is provided on the top of the staining plate, and the concentration indicator bar is divided into four areas from light to dark by gradient colors, and the four areas correspond to the four groups of the climbing grooves respectively.
[0014] Preferably, the connecting mechanism includes a card slot arranged on the outside of the dyeing plate, a card cover arranged on the bottom of the dyeing plate, and a silicone convex strip arranged on the inside of the card cover.
[0015] A cell slide immunofluorescence staining method, the specific steps are as follows:
[0016] S1: Use cell culture plates to make cell slides;
[0017] S2: First, drop an appropriate amount of labeled specific fluorescent antibody into the device, and transfer the prepared cell slide to the slide groove containing the hydrophobic material film;
[0018] S3: Use the snap cover to fix the staining plate on the cell culture plate;
[0019] S4: Use the card slot to cover the black light-shielding cover on the dyeing plate for light-shielding.
[0020] Preferably, the specific fluorescent antibody added in step S2 covers the cell slide to a certain height.
[0021] Compared with the prior art, the present invention has the following beneficial effects:
[0022] The present invention arranges four groups of conical slide grooves of different volumes on the staining plate, and personnel can select corresponding slide grooves according to the size of the cell slide and the concentration of the dyed antibody, so as to avoid antibody waste and facilitate personnel to take out the cell slide. A hydrophobic material film is arranged on the inner surface of the slide groove, which can effectively avoid the infiltration phenomenon caused by the hydrophilic material of the dyeing carrier, so that the specific fluorescent antibody is more evenly distributed on the cell slide, avoiding large differences in local dyeing effects caused by uneven antibody concentration, and at the same time reducing the total amount of specific fluorescent antibodies added, thereby reducing the test cost. BRIEF DESCRIPTION OF THE DRAWINGS
[0023] In order to more clearly illustrate the technical solutions of the embodiments of the present invention, the following briefly introduces the drawings required for describing the embodiments. Obviously, the drawings described below are only some embodiments of the present invention. For ordinary technicians in this field, other drawings can be obtained based on these drawings without creative labor. Among them:
[0024] Figure 1 It is a schematic diagram of the overall structure of an embodiment of the present invention;
[0025] Figure 2A bottom view of the present invention;
[0026] Figure 3 It is a schematic diagram of the internal structure of the present invention;
[0027] Figure 4 It is a schematic diagram of the overall structure of another embodiment of the present invention;
[0028] Figure 5 A flowchart of the method steps of the present invention;
[0029] Figure 6 It is a schematic diagram of the local structure of the sheet climbing groove of the present invention.
[0030] In the figure: 1. main body; 101. dyeing plate; 102. sheet climbing groove; 103. concentration indicator strip; 104. hydrophobic material film; 2. connecting mechanism; 201. card cover; 202. card slot; 203. silicone convex strip. DETAILED DESCRIPTION
[0031] In order to make the above-mentioned objects, features and advantages of the present invention more obvious and easy to understand, the specific implementation methods of the present invention are described in detail below in conjunction with the accompanying drawings.
[0032] In the following description, many specific details are set forth to facilitate a full understanding of the present invention, but the present invention may also be implemented in other ways different from those described herein, and those skilled in the art may make similar generalizations without violating the connotation of the present invention. Therefore, the present invention is not limited to the specific embodiments disclosed below.
[0033] Secondly, the term "one embodiment" or "embodiment" as used herein refers to a specific feature, structure, or characteristic that may be included in at least one implementation of the present invention. The term "in one embodiment" that appears in different places in this specification does not necessarily refer to the same embodiment, nor does it refer to a separate or selective embodiment that is mutually exclusive with other embodiments.
[0034] As attached Figure 1 To Attachment Figure 5 As shown:
[0035] Embodiment 1: This embodiment provides a cell climbing slice immunofluorescence staining device, including a main body 1, including a staining plate 101, four groups of climbing slice grooves 102 arranged on the staining plate 101, and a hydrophobic material film 104 arranged in the climbing slice grooves 102; a connecting mechanism 2, arranged on the staining plate 101, wherein the climbing slice grooves 102 are arranged on the upper surface of the staining plate 101.
[0036] The arrangement of the slide groove 102 can limit the position of the cell slide, avoid the slide shifting due to the vibration of the staining plate 101 during the light-proof incubation, and avoid the waste caused by uneven dye. The arrangement of four sets of slide grooves 102 can be selected according to the specifications of the cell slide to be stained, avoid the inconsistent specifications of the slide groove 102 and the cell slide, resulting in more waste of the subsequent added specific fluorescent antibodies, and then set a hydrophobic material film 104 in the slide groove 102. Since the glass surface has a strong hydrophilicity and a strong affinity for water droplets, when the water droplets fall on the glass surface, the intermolecular force on the glass surface makes the water molecules arrange more closely, so that The formation of a thin layer of water film and the phenomenon that water droplets expand outward on a flat surface such as glass is called infiltration. The infiltration phenomenon will affect the distribution of specific fluorescent antibodies, making the distribution of specific fluorescent antibodies on the cell slide uneven, resulting in some areas being overly stained, while other areas are stained differently, affecting the accuracy of subsequent personnel in observing and analyzing cell characteristics. The hydrophobic material film 104 can effectively avoid the infiltration phenomenon caused by the addition of specific fluorescent antibodies to the staining plate 101, making the specific fluorescent antibodies more evenly distributed on the cell slide, thereby reducing the total amount of specific fluorescent antibodies added, reducing the experimental cost, and reducing the background signal, and improving the specificity and contrast of staining.
[0037] Specifically, the volumes of the four groups of slice grooves 102 are different, namely 10 microliters, 20 microliters, 40 microliters and 80 microliters, and the diameters of the bottoms of the four groups of slice grooves 102 are 10 mm, 16 mm, 22 mm and 26 mm, respectively. When the cell slice is placed in the slice groove 102 for staining, there is a certain gap between the cell slice and the inner wall of the slice groove 102, which is convenient for the staff to take out the cell slice. The bottom surface of the slice groove 102 can also be set to an elliptical shape, with its short axis matching the cell slice, so as to limit the position of the cell slice and ensure stability during staining, while there is a certain gap between the long axis and the cell slice, which is convenient for the staff to take out the cell slice.
[0038] Specifically, the hydrophobic material film 104 is a PTFE film.
[0039] Specifically, the sheet climbing groove 102 is configured with a conical round bottom, with a large upper opening and a small lower round bottom.
[0040] Specifically, the hydrophobic material film 104 is located on the round bottom of the sheet climbing groove 102 .
[0041] Specifically, a concentration indicator bar 103 is provided on the top of the staining plate 101 . The concentration indicator bar 103 is divided into four areas from light to dark by gradient colors. The four areas correspond to the four groups of sheet climbing grooves 102 , respectively.
[0042] Specifically, the connecting mechanism 2 includes a card slot 202 arranged on the outside of the dyeing plate 101 , a card cover 201 arranged on the bottom of the dyeing plate 101 , and a silicone convex strip 203 arranged on the inside of the card cover 201 .
[0043] As can be seen from the above, during the staining work, the cell slide is placed in the slide groove 102, and then the specific fluorescent antibody is added into the slide groove 102 to stain the cell slide. The conical round bottom slide groove 102 with a large upper opening and a small lower opening can make the added specific fluorescent antibody completely act on the cell slide, and it is convenient for personnel to take out the cell slide later;
[0044] By setting the hydrophobic material film 104 on the round bottom of the cell-climbing groove 102, the specific fluorescent antibody can be prevented from diffusing at the bottom, so that the antibody is more concentrated, which helps the antibody to effectively bind to the cell-climbing slide, reduces the amount of antibody used, and can better remove unbound or non-specifically bound antibodies, thereby improving the specificity of the fluorescent signal and allowing the specific fluorescent antibody to flow relatively freely on the inner surface of the groove, which can promote the attachment and growth of cells to the surface, which is crucial to the effectiveness of cell culture and experimental results;
[0045] Four groups of slice grooves 102 with different volumes are set, and concentration indicator bars 103 are set to indicate the slice grooves 102 with different volumes, so that when personnel select the slice groove 102 for staining experiments, they can easily know the concentration of specific fluorescent antibodies when selecting the slice groove 102 for the experiment. After placing the cell slice on the staining plate 101, the staining plate 101 is clamped on the cell culture plate for making the slice through the card cover 201, and the silicone convex strips 203 will fill the gap between the card cover 201 and the culture plate to improve the clamping stability, and then the card slot 202 will clamp the opaque light-shielding cover on the staining plate 101 to prevent the dye from evaporating.
[0046] Embodiment 2: This embodiment provides a cell climbing slice immunofluorescence staining device, including a main body 1, including a staining plate 101, four groups of climbing slice grooves 102 arranged on the staining plate 101, and a hydrophobic material film 104 arranged in the climbing slice grooves 102; a connecting mechanism 2, which is arranged on the staining plate 101.
[0047] Specifically, the volumes of the four groups of slice climbing grooves 102 are different, namely 10 microliters, 20 microliters, 40 microliters and 80 microliters, and the diameters of the bottoms of the four groups of slice climbing grooves 102 are 10 mm, 16 mm, 22 mm and 26 mm, respectively.
[0048] Specifically, the hydrophobic material film 104 is a PTFE film.
[0049] Specifically, the sheet climbing groove 102 is configured with a conical round bottom, with a large upper opening and a small lower round bottom.
[0050] Specifically, the hydrophobic material film 104 is located on the round bottom of the sheet climbing groove 102 .
[0051] Specifically, the hydrophobic material film 104 is located on the conical inner wall of the sheet climbing groove 102 .
[0052] Specifically, a concentration indicator bar 103 is provided on the top of the staining plate 101 . The concentration indicator bar 103 is divided into four areas from light to dark by gradient colors. The four areas correspond to the four groups of sheet climbing grooves 102 , respectively.
[0053] Specifically, the connecting mechanism 2 includes a card slot 202 arranged on the outside of the dyeing plate 101 , a card cover 201 arranged on the bottom of the dyeing plate 101 , and a silicone convex strip 203 arranged on the inside of the card cover 201 .
[0054] As can be seen from the above, during the staining work, the cell slide is placed in the slide groove 102, and then the specific fluorescent antibody is added into the slide groove 102 to stain the cell slide. The conical round bottom slide groove 102 with a large upper opening and a small lower opening can make the added specific fluorescent antibody completely act on the cell slide, and it is convenient for personnel to take out the cell slide later;
[0055] By arranging the hydrophobic material film 104 on the round bottom of the climbing slice groove 102, the specific fluorescent antibody can be prevented from diffusing at the bottom, so that the antibody is more concentrated, which is helpful for the antibody to effectively bind to the cell climbing slice, so that the cell climbing slice is more fully in contact with the dye, and the amount of antibody used is reduced, and the unbound or non-specifically bound antibodies can be better removed, thereby improving the specificity of the fluorescent signal. The hydrophobic material film 104 is also arranged on the conical inner wall of the climbing slice groove 102, which can prevent the infiltration phenomenon to the greatest extent, and facilitate personnel to more comprehensively control the behavior of the antibody. At the same time, it can reduce the difference between samples caused by different surface hydrophilicity, improve the repeatability and consistency of the experiment, and reduce the influence of unnecessary surface tension on cells, thereby protecting the cell structure;
[0056] Four groups of slice climbing grooves 102 with different volumes are set, and concentration indicator bars 103 are set to indicate the slice climbing grooves 102 with different volumes, so that when personnel select the slice climbing groove 102 for staining experiments, they can easily know the specific fluorescent antibody concentration when selecting the slice climbing groove 102 for the experiment. After placing the cell slice on the staining plate 101, the staining plate 101 is clamped on the cell culture plate for making the slice through the card cover 201. The silicone convex strip 203 will fill the gap between the card cover 201 and the culture plate to improve the clamping stability. Then the card slot 202 will clamp the opaque light-shielding cover on the staining plate 101 to prevent the dye from evaporating. When the work is completed, the card slot 202 can also be used to clamp the cover of the culture plate on the staining plate 101 to reduce the occupied space.
[0057] Embodiment 3: This embodiment is basically the same as the previous embodiment, except that the main body 1 is made of PTFE hydrophobic material as a whole, and no hydrophobic material film 104 is required.
[0058] Embodiment 4: This embodiment provides a method for immunofluorescence staining of cell slides, and the specific steps are as follows:
[0059] S1: Use cell culture plates to make cell slides;
[0060] S2: First, drop a proper amount of labeled specific fluorescent antibody into the device, and transfer the prepared cell slide to the slide groove 102 to which the hydrophobic material film is adhered;
[0061] S3: Use the card cover 201 to buckle the staining plate 101 onto the cell culture plate and fix it;
[0062] S4: Use the card slot 202 to cover the black light-shielding cover on the dyeing plate 101 to perform light-shielding dyeing.
[0063] Specifically, the specific fluorescent antibody added in step S2 covers the cell slide to a certain height.
[0064] As attached Figure 6 As shown:
[0065] Embodiment 5: This embodiment is basically the same as the previous embodiment, except that the climbing slice groove 102 on the staining plate 101 is set to be through-through from top to bottom, and the hydrophobic material film 104 is located at the bottom opening of the climbing slice groove 102 and has a certain elasticity. During the staining work, the card cover 201 and the silicone convex strip 203 are used to clamp the device on the horizontal plate, which not only has the effect of reinforcing the device as a whole, but also supports the water transfer material film 104 to prevent it from being deformed due to its own elasticity and the squeezing of specific fluorescent antibodies and cell climbing slices, thereby affecting the staining effect. After the staining work is completed, the horizontal plate is removed, and then the water transfer material film 104 is lifted up. It will bulge upward using its own elasticity, and then lift up the cell climbing slice inside it, making it convenient for personnel to remove the cell climbing slice.
[0066] Importantly, it should be noted that the construction and arrangement of the present application shown in a plurality of different exemplary embodiments are only exemplary. Although only a few embodiments are described in detail in this disclosure, it should be readily understood by those who refer to this disclosure that many modifications are possible (e.g., the size, scale, structure, shape and proportion of various elements, and parameter values (e.g., temperature, pressure, etc.), installation arrangement, use of materials, color, directional changes, etc.) without substantially departing from the novel teachings and advantages of the subject matter described in the application. For example, the element shown as integrally formed can be composed of multiple parts or elements, the position of the element can be inverted or otherwise changed, and the nature or number or position of the discrete element can be changed or changed. Therefore, all such modifications are intended to be included in the scope of the present invention. The order or sequence of any process or method steps can be changed or reordered according to alternative embodiments. In the claims, any "device plus function" clause is intended to cover the structure of performing the function described herein, and is not only structurally equivalent but also equivalent structure. Without departing from the scope of the present invention, other replacements, modifications, changes and omissions can be made in the design, operating conditions and arrangement of the exemplary embodiments. Therefore, the invention is not limited to a specific embodiment, but extends to numerous modifications still falling within the scope of the appended claims.
[0067] Additionally, in order to provide a concise description of exemplary embodiments, all features of an actual embodiment (ie, those features that are not relevant to the best mode presently contemplated for carrying out the invention or those that are not relevant to implementing the invention) may not be described.
[0068] It will be appreciated that in the development of any actual implementation, as in any engineering or design project, numerous implementation-specific decisions may be made. Such a development effort may be complex and time-consuming, but will be a routine task of design, fabrication, and production for those of ordinary skill having the benefit of this disclosure without undue experimentation.
[0069] It should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention rather than to limit it. Although the present invention has been described in detail with reference to the preferred embodiments, those skilled in the art should understand that the technical solutions of the present invention may be modified or replaced by equivalents without departing from the spirit and scope of the technical solutions of the present invention, which should all be included in the scope of the claims of the present invention.
Claims
1. A cell slide immunofluorescence staining device, characterized in that: include, The main body (1) comprises a dyeing plate (101), four groups of sheet climbing grooves (102) arranged on the dyeing plate (101), and a hydrophobic material film (104) arranged in the sheet climbing grooves (102); The connecting mechanism (2) is arranged on the dyeing plate (101).
2. A cell slide immunofluorescence staining device according to claim 1, characterized in that: The volumes of the four groups of the climbing slice grooves (102) are different, namely 10 microliters, 20 microliters, 40 microliters and 80 microliters, and the diameters of the bottoms of the four groups of the climbing slice grooves (102) are 10 mm, 16 mm, 22 mm and 26 mm, respectively.
3. A cell slide immunofluorescence staining device according to claim 1, characterized in that: The hydrophobic material film (104) is a PTFE film.
4. The cell slide immunofluorescence staining device according to claim 1, characterized in that: The sheet climbing groove (102) is configured with a conical round bottom, with a large upper opening and a small lower round bottom.
5. A cell slide immunofluorescence staining device according to claim 4, characterized in that: The hydrophobic material film (104) is located on the round bottom of the sheet climbing groove (102).
6. A cell slide immunofluorescence staining device according to claim 4, characterized in that: The hydrophobic material film (104) is located on the conical inner wall of the sheet climbing groove (102).
7. The cell slide immunofluorescence staining device according to claim 1, characterized in that: A concentration indicator bar (103) is provided on the top of the dyeing plate (101), and the concentration indicator bar (103) is divided into four areas from light to dark by gradient color, and the four areas correspond to the four groups of the sheet climbing grooves (102) respectively.
8. The cell slide immunofluorescence staining device according to claim 1, characterized in that: The connecting mechanism (2) comprises a card slot (202) arranged on the outside of the dyeing plate (101), a card cover (201) arranged on the bottom of the dyeing plate (101), and a silicone convex strip (203) arranged on the inside of the card cover (201).
9. A cell slide immunofluorescence staining method, characterized in that: The specific steps are as follows: S1: Use cell culture plates to make cell slides; S2: First, drop a proper amount of labeled specific fluorescent antibody into the device, and transfer the prepared cell slide to the slide groove (102) containing the hydrophobic material film; S3: Using the card cover (201) to buckle the staining plate (101) onto the cell culture plate and fix it; S4: Using the card slot (202) to cover the black light-shielding cover plate on the dyeing plate (101) to perform light-shielding dyeing.
10. A cell slide immunofluorescence staining method according to claim 9, characterized in that: The specific fluorescent antibody added in step S2 is not higher than a certain height of the cell slide.