A spotting device for protein dot blotting experiments

Abstract

This utility model relates to the field of biological experimental equipment technology. It discloses a sample application device for protein dot blot experiments, comprising a base and a cover plate. The cover plate has a first side and a second side facing each other, with the second side of the cover plate facing the base. The first side of the cover plate has a plurality of sample application holes arranged in an array at intervals, penetrating the cover plate and extending to the second side of the cover plate. A membrane is placed between the base and the second side of the cover plate. This application, by using a cover plate with a plurality of sample application holes covering the membrane, ensures the standardization of the sample application position, allowing each sample to form an independent diffusion area, effectively avoiding physical mixing and cross-contamination between samples, and ensuring the accuracy of experimental results. Furthermore, the arrayed sample application holes achieve a consistent sample application distance, which helps image analysis software accurately identify and locate each sample point, improving the accuracy and efficiency of data analysis and reducing manual correction operations.

Description

Technical Field

[0001] This utility model relates to the field of biological experimental equipment technology, and in particular to a spotting device for protein dot blot experiments. Background Technology

[0002] Western blot assay is a commonly used protein detection technique. This technique involves spotting protein samples onto a membrane sheet, followed by detection using specific antibodies, thereby achieving qualitative or quantitative analysis of specific proteins. Spotting is a crucial step in this technique, as its quality directly affects the accuracy and reliability of the experimental results.

[0003] Currently, in spot experiments, samples are absorbed on the membrane and then diffuse. If the spotting distance is too close, the diffusion areas of adjacent samples may overlap, causing cross-contamination and false positive signals, thus affecting the accuracy of the experimental results. Furthermore, inconsistent spotting distances can also affect the accurate identification and positioning by image analysis software, leading to data analysis bias. This necessitates additional image calibration and manual segmentation of overlapping spots, reducing experimental efficiency and increasing costs. Utility Model Content

[0004] To address the aforementioned technical problems, this invention proposes a spotting device for protein dot blot experiments, which can avoid physical mixing and cross-contamination between samples, and helps image analysis software accurately identify and locate each sample spot, thereby improving experimental efficiency and reducing experimental costs.

[0005] To achieve the above objectives, this utility model provides a sample application device for a protein dot blot experiment, comprising a base and a cover plate. The cover plate has a first side and a second side facing each other, with the second side of the cover plate facing the base. The first side of the cover plate is provided with a plurality of sample application holes arranged in a spaced array, the sample application holes penetrating the cover plate and extending to the second side of the cover plate. A membrane is disposed between the base and the second side of the cover plate.

[0006] In one alternative embodiment, the base is provided with a limiting structure adapted to restrict the cover plate from moving horizontally.

[0007] In one alternative embodiment, the limiting structure is a receiving groove provided on the base, the second side of the cover plate is disposed in the receiving groove, and the outer side wall of the cover plate is adapted to the inner side wall of the receiving groove.

[0008] In one optional embodiment, the plurality of sample dispensing wells are configured as a plurality of first sample dispensing wells and a plurality of second sample dispensing wells, wherein the plurality of first sample dispensing wells are arranged in an array, and the plurality of second sample dispensing wells are arranged in an array; wherein.

[0009] The diameter of the first sample feeding port is smaller than the diameter of the second sample feeding port;

[0010] And / or, the distance between two adjacent first sample feeding holes is less than the distance between two adjacent second sample feeding holes.

[0011] In one optional embodiment, the first side of the cover plate is provided with two aperture indication marks, which are respectively provided with the first sample feeding hole and the second sample feeding hole.

[0012] In one alternative embodiment, a counting meter is provided on the first side of the cover plate, and the counting meter is provided corresponding to the sample dispensing hole.

[0013] In one alternative embodiment, the first side of the cover plate is further provided with a mounting hole, and a handle is detachably connected to the mounting hole.

[0014] In one alternative embodiment, a connecting structure is provided between the cover plate and the base, and the cover plate is rotatably connected to the base through the connecting structure.

[0015] In one alternative embodiment, the connection structure includes a connecting seat and a pin, the connecting seat being connected to the side wall of the cover plate, the pin being connected to one of the base and the connecting seat, and the pin being rotatably connected to the other of the base and the connecting seat.

[0016] In one alternative embodiment, the side wall of the base has a notch that communicates with the receiving groove.

[0017] Compared with the prior art, the present invention has the following advantages and technical effects:

[0018] 1. By setting a cover plate with several sample application holes to cover the membrane, the standardization of the sample application position is ensured, so that each sample forms an independent diffusion area, effectively avoiding physical mixing and cross-contamination between samples, and ensuring the accuracy of experimental results.

[0019] 2. The array-set sample wells achieve a consistent sampling distance, which helps image analysis software accurately identify and locate each sample point, improving the accuracy and efficiency of data analysis and reducing manual calibration operations.

[0020] 3. By pre-planning the sampling locations and distances, random sampling is avoided, the effective area of ​​the membrane is fully utilized, the sample detection throughput of a single experiment is increased, reagent waste caused by uneven sample distribution is avoided, experimental efficiency is improved, and experimental costs are reduced. Attached Figure Description

[0021] The accompanying drawings, which form part of this application, are used to provide a further understanding of this application. The illustrative embodiments and descriptions of this application are used to explain this application and do not constitute an undue limitation of this application. In the drawings:

[0022] Figure 1 This is a schematic diagram of the structure with the cover plate closed.

[0023] Figure 2 A schematic diagram of the structure with the cover plate in the open position;

[0024] Figure 3 This is a schematic diagram of the cover plate.

[0025] Figure 4 This is a schematic diagram of the base structure;

[0026] Figure 5 This is a structural diagram of the handle;

[0027] Figure 6 This is a structural diagram of the pin.

[0028] Figure 7 This is a schematic diagram illustrating the positional relationship between the notch and the base.

[0029] Figure 8 This is a schematic diagram illustrating another positional relationship between the notch and the base;

[0030] Figure 9 This is a schematic diagram showing the positional relationship between the insertion column and the sample application hole.

[0031] The components are as follows: 1. Base; 2. Cover plate; 201. First side; 202. Second side; 3. Receiving groove; 4. First sample feeding hole; 5. Second sample feeding hole; 6. Hole diameter indication mark; 7. Counting meter; 8. Mounting hole; 9. Handle; 10. Connecting structure; 101. Connecting seat; 102. Pin; 11. Notch; 12. Cover plate; 13. Insert post. Detailed Implementation

[0032] The technical solutions of the present utility model will be clearly and completely described below with reference to the accompanying drawings of the embodiments. Obviously, the described embodiments are only some embodiments of the present utility model, and not all embodiments. Based on the embodiments of the present utility model, all other embodiments obtained by those of ordinary skill in the art without creative effort are within the protection scope of the present utility model.

[0033] To make the above-mentioned objectives, features and advantages of this utility model more apparent and understandable, the utility model will be further described in detail below with reference to the accompanying drawings and specific embodiments.

[0034] Reference Figures 1-9This invention provides a sample application device for a protein dot blot experiment, including a base 1 and a cover plate 2. The cover plate 2 has a first side 201 and a second side 202 facing each other. The second side 202 of the cover plate 2 faces the base 1. The first side 201 of the cover plate 2 is provided with a plurality of sample application holes arranged in an array at intervals. The sample application holes penetrate the cover plate 2 and extend to the second side 202 of the cover plate 2. A membrane is placed between the base 1 and the second side 202 of the cover plate 2.

[0035] In this embodiment, the second side 202 of the base 1 and the cover plate 2 cooperate to form a clamping structure. During the sample dispensing operation, the membrane is clamped between the base 1 and the second side 202 of the cover plate 2. The second side 202 of the base 1 and the cover plate 2 are used to clamp and fix the membrane, preventing relative movement of the membrane during the sample dispensing operation, thereby improving experimental accuracy. Furthermore, along the height direction of the sample dispensing device, the cover plate 2 is preferably positioned above the base 1. This facilitates the sample dispensing operation and also makes it easier to separate the cover plate 2 from the base 1, thus facilitating the removal of the membrane.

[0036] In this embodiment, a plurality of sample application holes are provided on the first side 201 of the cover plate 2. The sample application holes penetrate the cover plate 2 and correspond to the membrane on the base 1. The plurality of sample application holes are arranged at intervals, so that there is a preset distance between adjacent sample application holes, thereby enabling each sample to form an independent diffusion area, effectively avoiding physical mixing and cross-contamination between samples, and ensuring the accuracy of experimental results. Moreover, the array arrangement of the plurality of sample application holes ensures that the plurality of sample application holes achieve a consistent sampling distance, which helps the image analysis software to accurately identify and locate each sample point, improves the accuracy and efficiency of data analysis, and reduces manual correction operations.

[0037] Optionally, the membrane described above can be made of PVDF. Of course, other types of membrane materials can also be selected according to the actual experimental needs.

[0038] In one embodiment, a limiting structure is provided on the base 1, which is adapted to restrict the cover plate 2 from moving in the horizontal direction.

[0039] In this embodiment, after the cover plate 2 and the base 1 clamp the membrane, the limiting structure restricts the movement of the cover plate 2 relative to the base 1, so as to avoid the flatness of the membrane being affected by the movement of the cover plate 2, improve the accuracy of the spotting, thereby avoiding physical mixing and cross-contamination between samples, and helping to accurately identify and locate each sample point in the later stage through image analysis software.

[0040] Optionally, the limiting structure can be made of bolted connectors, magnetic connectors or snap-fit ​​connectors, so that the cover plate 2 can be detachably connected to the base 1 by bolts, magnetic attraction or snap-fit, so that the cover plate 2 can be fixed relative to the base 1 during the sampling operation and can be separated from the base 1 after the sampling is completed.

[0041] Furthermore, referring to Figure 9 The device is also equipped with a cover plate 12, on one end face of which are fixed several inserts 13, each insert 13 corresponding to a sample dispensing hole. Thus, when storing or cleaning the dispensing device, the inserts 13 can be inserted into each sample dispensing hole to seal or clean them, facilitating storage and cleaning and improving the device's practicality.

[0042] In one embodiment, refer to Figure 2 , Figure 4 The limiting structure is a receiving groove 3 set on the base 1, the second side 202 of the cover plate 2 is set in the receiving groove 3, and the outer side wall of the cover plate 2 is adapted to the inner side wall of the receiving groove 3.

[0043] In this embodiment, the cover plate 2 is detachably connected to the base 1 via a snap-fit ​​mechanism. When clamping the membrane, the second side 202 of the cover plate 2 is placed into the receiving groove 3. The side wall of the receiving groove 3 limits the cover plate 2, preventing it from moving horizontally. After the sample is applied, the cover plate 2 can be lifted vertically to separate it from the base 1. This allows the cover plate 2 to be stably placed on the base 1, preventing horizontal displacement during use and ensuring precise alignment between the sample application hole and the target position on the membrane, thereby improving the accuracy of the sample application. Furthermore, the snap-fit ​​mechanism eliminates the need for additional limiting devices, simplifying the overall design of the sample application device and reducing operating costs, thus enhancing its practicality.

[0044] Furthermore, when the cover plate 2 is located in the receiving groove 3, the first side 201 of the cover plate 2 is preferably located outside the receiving groove 3, so that the top of the cover plate 2 protrudes out of the receiving groove 3, which facilitates the subsequent separation of the cover plate 2 from the base 1.

[0045] In one embodiment, refer to Figure 1 , Figure 2 , Figure 3 The sample feeding holes are configured as multiple first sample feeding holes 4 and multiple second sample feeding holes 5, the multiple first sample feeding holes 4 are arranged in an array, and the multiple second sample feeding holes 5 are arranged in an array; wherein, the diameter of the first sample feeding hole 4 is smaller than the diameter of the second sample feeding hole 5; and / or, the distance between two adjacent first sample feeding holes 4 is smaller than the distance between two adjacent second sample feeding holes 5.

[0046] In this embodiment, to adapt to different experimental needs, sample dispensing holes with different apertures and different adjacent spacings can be provided on the cover plate 2. For example, the aperture of the first sample dispensing hole 4 is smaller than that of the second sample dispensing hole 5, and the spacing between two adjacent first sample dispensing holes 4 is smaller than the spacing between two adjacent second sample dispensing holes 5. In this way, the first sample dispensing hole 4 is suitable for precise spotting of small-volume samples, and the second sample dispensing hole 5 is suitable for spotting large-volume samples, improving the applicability and flexibility of the spotting device.

[0047] Of course, depending on the experimental requirements, the diameter of the first sample feeding hole 4 can be set to be different from that of the second sample feeding hole 5, and the distance between two adjacent first sample feeding holes 4 can be the same as the distance between two adjacent second sample feeding holes 5. Alternatively, the diameter of the first sample feeding hole 4 can be set to be the same as that of the second sample feeding hole 5, and the distance between two adjacent first sample feeding holes 4 can be different from the distance between two adjacent second sample feeding holes 5.

[0048] Of course, in addition to the two types of sample wells mentioned above, various other sample well models can be set to meet different experimental needs.

[0049] In one embodiment, refer to Figure 1 , Figure 2 , Figure 3 The first side 201 of the cover plate 2 is provided with two aperture indication marks 6, which correspond to the first sample feeding hole 4 and the second sample feeding hole 5 respectively.

[0050] In this embodiment, by setting the aperture indication mark 6, the experimenter can more intuitively distinguish between different types of sample loading wells, avoid confusion during the experiment, and improve experimental efficiency and accuracy.

[0051] In one embodiment, a counting meter 7 is provided on the first side 201 of the cover plate 2, and the counting meter 7 is provided corresponding to the sample dispensing hole.

[0052] In this embodiment, the setting of the counting table 7 makes it easy for the experimenter to record and track the well positions that have been sampled, avoiding repeated sampling or omissions. Especially when sampling a large number of samples, it can effectively improve work efficiency and accuracy.

[0053] Specifically, when the sample wells are arranged in an array, the counting table 7 can be formed by a combination of horizontal and vertical counting, which makes it easier for experimenters to visually distinguish sample wells in different positions, avoid confusion during the experiment, and improve experimental efficiency and accuracy.

[0054] In one embodiment, refer to Figure 1 , Figure 3 , Figure 5 , Figure 6 The first side 201 of the cover plate 2 is also provided with a mounting hole 8, and a handle 9 is detachably connected to the mounting hole 8.

[0055] In this embodiment, a handle 9 is provided on the first side 201 of the cover plate 2, which facilitates the movement of the cover plate 2 and makes it easy to separate the cover plate 2 from the base 1.

[0056] In one embodiment, refer to Figure 1 , Figure 2A connecting structure 10 is provided between the cover plate 2 and the base 1, and the cover plate 2 is rotatably connected to the base 1 through the connecting structure 10.

[0057] In this embodiment, the cover plate 2 and the base 1 are rotatably connected by the connecting structure 10, thereby preventing the cover plate 2 from separating from the base 1, facilitating the replacement or adjustment of the membrane, and also making it convenient to store the sample dispensing device.

[0058] In one embodiment, refer to Figure 1 , Figure 2 The connecting structure 10 includes a connecting seat 101 and a pin 102. The connecting seat 101 is connected to the side wall of the cover plate 2, and the pin 102 is connected to one of the base 1 and the connecting seat 101. The pin 102 is rotatably connected to the other of the base 1 and the connecting seat 101.

[0059] In this embodiment, the pin 102 is connected to the base 1 and the pin 102 is rotatably connected to the connecting seat 101 as an example. One end of the pin 102 is fixed to the base 1, and the other end of the pin 102 passes through the connecting seat 101 and is rotatably connected to the connecting seat 101, so that the cover plate 2 can rotate relative to the base 1. This makes it convenient for the experimenter to replace or adjust the diaphragm without completely separating the cover plate 2 and the base 1, thus improving the convenience and efficiency of operation.

[0060] In one embodiment, refer to Figure 7 , Figure 8 The side wall of the base 1 has a notch 11, which is connected to the receiving groove 3.

[0061] In this embodiment, a receiving groove 3 is provided on the base 1. In order to facilitate the removal of the membrane from the groove after the sample is applied, a notch 11 is provided to facilitate the operation of the experimenter and improve the ease of use of the device.

[0062] The notch 11 can be provided in one location on the side wall of the base 1 away from the connecting structure 10. Of course, in addition to the side wall of the base 1 having the connecting structure 10, multiple notches 11 can also be provided on other side walls of the base 1.

[0063] In one specific embodiment, the membrane is immersed in methanol for 5 minutes for permeation treatment to enhance its protein binding capacity. The pretreated membrane is then placed flat on base 1, ensuring it is wrinkle-free. A 45° angle cut (approximately 2 mm) is used as an orientation reference mark to prevent membrane misalignment during subsequent operations.

[0064] Then, cover the membrane with the pre-set hole position cover plate 2 and align it to ensure that the center of the sample loading hole corresponds precisely to the target sample loading position.

[0065] Next, use a pipette to draw 5 μL or 10 μL of sample (select a fixed volume according to experimental needs; this sample application device can simultaneously meet different sample volume requirements), avoiding the formation of air bubbles. Keep the pipette perpendicular to the membrane surface, lightly touch the center of the well with the pipette tip, and slowly push out the sample, ensuring that each sample point has a consistent volume and no splashing.

[0066] Spot the samples one by one in the preset order (e.g., from left to right, from top to bottom) to avoid missing or repeating spots. After spotting, place the entire spotting device in a 45°C oven and dry for 10 minutes to promote strong protein binding to the membrane surface. Then open cover 2, confirm the orientation according to the cut corner mark in the upper left corner, transfer the membrane to the antibody incubation box, and perform subsequent operations such as blocking, primary antibody incubation, and secondary antibody incubation according to the experimental procedure.

[0067] The above are merely preferred embodiments of this application, but the scope of protection of this application is not limited thereto. Any variations or substitutions that can be easily conceived by those skilled in the art within the scope of the technology disclosed in this application should be included within the scope of protection of this application. Therefore, the scope of protection of this application should be determined by the scope of the claims.

Claims

1. A spotting apparatus for protein dot blot assay, characterized in that: include: Base (1); The cover plate (2) has a first side (201) and a second side (202) facing each other. The second side (202) of the cover plate (2) faces the base (1). The first side (201) of the cover plate (2) is provided with a plurality of sample holes in a spaced array. The sample holes penetrate the cover plate (2) and extend to the second side (202) of the cover plate (2). A diaphragm is disposed between the base (1) and the second side (202) of the cover plate (2).

2. The spotting apparatus for protein dot blot assay according to claim 1, characterized in that: The base (1) is provided with a limiting structure, which is adapted to restrict the cover plate (2) from moving in the horizontal direction.

3. The spotting apparatus for protein dot blot assay according to claim 2, characterized in that: The limiting structure is a receiving groove (3) provided on the base (1), the second side (202) of the cover plate (2) is provided in the receiving groove (3), and the outer side wall of the cover plate (2) is adapted to the inner side wall of the receiving groove (3).

4. The spotting apparatus for protein dot blot assay according to claim 1, characterized in that: The plurality of sample dispensing holes are configured as a plurality of first sample dispensing holes (4) and a plurality of second sample dispensing holes (5), wherein the plurality of first sample dispensing holes (4) are arranged in an array and the plurality of second sample dispensing holes (5) are arranged in an array; wherein, The diameter of the first sample feeding hole (4) is smaller than the diameter of the second sample feeding hole (5); And / or, the distance between two adjacent first sample feeding holes (4) is less than the distance between two adjacent second sample feeding holes (5).

5. The spotting apparatus for protein dot blot assay according to claim 4, characterized in that: The cover plate (2) has two aperture markings (6) on its first side (201), and the two aperture markings (6) are respectively set to correspond to the first sample feeding hole (4) and the second sample feeding hole (5).

6. The spotting apparatus for protein dot blot assay according to claim 1, characterized in that: A counting table (7) is provided on the first side (201) of the cover plate (2), and the counting table (7) is provided in correspondence with the sample dispensing hole.

7. The spotting apparatus for protein dot blot assay according to any one of claims 1-6, characterized in that: The cover plate (2) is also provided with a mounting hole (8) on the first side (201), and a handle (9) is detachably connected to the mounting hole (8).

8. The spotting apparatus for protein dot blot assay according to any one of claims 1-6, characterized in that: A connecting structure (10) is provided between the cover plate (2) and the base (1), and the cover plate (2) is rotatably connected to the base (1) through the connecting structure (10).

9. The spotting apparatus for protein dot blot assay according to claim 8, characterized in that: The connection structure (10) includes: The connecting seat (101) is connected to the side wall of the cover plate (2); A pin (102) is connected to one of the base (1) and the connecting seat (101), and the pin (102) is rotatably connected to the other of the base (1) and the connecting seat (101).

10. The spotting apparatus for protein dot blot assay according to claim 3, characterized in that: The base (1) has a notch (11) on its side wall, and the notch (11) is connected to the receiving groove (3).

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