Improved perforated plate convenient for serum multi-factor mixed detection

Abstract

The utility model relates to an improved perforated plate convenient for serum multi-factor mixed detection, which comprises a plate body, a plurality of main detection holes are arranged on the surface of the plate body, a plurality of transverse partition plates and longitudinal partition plates are arranged in the main detection holes, the transverse partition plates and the longitudinal partition plates are respectively connected with the inner walls of the main detection holes, and the heights of the transverse partition plates and the longitudinal partition plates are lower than the depth of the main detection holes. The transverse partition plates and the longitudinal partition plates divide the main detection hole into a plurality of sub detection holes with the depth smaller than that of the main detection hole. According to the improved perforated plate convenient for serum multi-factor mixed detection, liquid can be added into the plurality of sub-detection holes at one time so as to carry out multi-factor detection by using conventional equipment, so that the multi-factor detection cost and complexity are reduced.

Description

Technical Field

[0001] The utility model relates to a multi-hole plate used for sample index detection, in particular to an improved detection hole plate which is convenient for multi-factor mixed detection. Background Art

[0002] Serum (or in vitro culture fluid) cytokine detection is a very important part of modern medical research and clinical treatment. The difficulty in detecting cytokines in serum or culture fluid is that the sampling is small, while there are many cytokines to be tested. Therefore, the current common method is to use high-throughput multi-well plates for detection. Although this type of technology has greatly accelerated the process of multi-factor detection, the above two difficulties, namely, small sampling and many factors, still require operators to spend a lot of manpower, material resources and time, and have great uncontrollability in the repeatability and batch variability of the results. For this reason, a few companies have developed "microwell mixing" test methods or "microsphere mixing" detection methods to solve the problem of "multi-factor mixing" detection. At present, there are mainly the following multi-factor detection technology platforms on the market: 1) MSD technology platform, which is based on the principle of electrochemiluminescence and uses dot matrix technology to achieve simultaneous detection of multiple factors in 96-well graphite electrode plates, 2) Luminex platform technology, also known as "liquid suspension chip technology" in China. It organically integrates a number of the latest scientific and technological achievements, such as fluorescent coded microsphere technology, laser analysis technology, flow cytometry technology, high-speed digital signal processing technology, and computer algorithms. Using the principles of microspheres and flow cytometry, the microspheres contain three types of immunofluorescence, and 100 different microspheres can be distinguished by different ratios of fluorescence. Therefore, using microsphere technology, up to 100 proteins or genes can be detected simultaneously. 3) CBA platform technology, also known as cytokine microsphere detection technology. Micro-sample multi-index flow protein quantification technology is a multiple protein quantification detection method based on a flow cytometric detection system. The principle is similar to that of ELISA detection, that is, using tiny, dispersed particles to capture liquid analytes, and using flow cytometry to detect the fluorescence emitted by the "sandwich"-like particles-the analyte complex, thereby determining the number of analytes and achieving the purpose of detecting multiple factors at one time.

[0003] These mainstream technology platforms have undoubtedly brought great convenience to solving high-throughput sample multi-factor detection. However, the use of such technologies requires the purchase of special consumables and instruments. However, such consumables and instruments are not applicable to the microplates and plate readers currently used in laboratories, which makes it difficult to promote their use. Due to their high degree of professionalism and matching, these technologies are relatively complex in terms of detection consumables, personnel operation, result recording instruments, result analysis software, etc., and require more cost and energy investment. For general scientific researchers, this detection technology is difficult to implement in the absence of sufficient professionalism and funding.

[0004] Therefore, how to use existing standard consumables and instruments to achieve the same "multi-factor" and "mixed" detection purposes has become a technical "bridge" problem that urgently needs to be solved in the field of "multi-factor detection". A Chinese patent with the publication number "CN207780275 U" and the name "An ELISA sample loading device" discloses an ELISA plate body, and its ELISA holes are all grooves with uniform depth and flush surface. When using a traditional ELISA instrument for multi-factor detection, since the depth of each hole is consistent and the caliber of the hole itself is small, when inputting a liquid sample, the operator or manipulator needs to accurately inject the sample into each hole to maintain the consistency of the sample injection in each hole, which is extremely cumbersome to operate. Utility Model Content

[0005] In order to solve the technical problems of inconvenient and non-standard input of sample liquid and subsequent operations during the above-mentioned multi-factor mixed detection, the utility model provides a multi-factor mixed detection that can be performed using existing standard manual equipment or mechanical workbenches, and the results can be used to detect common signals using a standard plate reader, thereby facilitating laboratories and inspection departments with basic laboratory equipment to carry out multi-factor mixed detection, greatly reducing the technical difficulty and cost of multi-factor detection, and improving the detection speed, thereby facilitating an improved multi-well plate for serum multi-factor mixed detection.

[0006] The utility model is an improved multi-porous plate for facilitating serum multi-factor mixed detection, comprising a plate body, a plurality of main detection holes on the surface of the plate body, a plurality of transverse partitions and longitudinal partitions in the main detection hole, the transverse partitions and longitudinal partitions are respectively connected to the inner wall of the main detection hole, the height of the transverse partitions and longitudinal partitions is lower than the depth of the main detection hole, and the transverse partitions and longitudinal partitions divide the main detection hole into a plurality of sub-detection holes with a depth less than the main detection hole.

[0007] The advantage of the improved detection well plate for multi-factor mixed detection is that its main detection well is divided into a number of sub-detection wells with a depth lower than the main detection well by a horizontal partition and a longitudinal partition with a lower height in the well. In this way, when performing multi-factor detection, the operator only needs to add samples to the main detection well (such as using an 8-tube sampler commonly used in laboratories and adding samples according to a 96-well plate structure). As long as the height of the sample liquid in the main detection well is higher than the depth of the sub-detection well, the mixed sample can be evenly injected into each sub-detection well, which is very convenient to operate. Since primary antibodies targeting different factors have been added to each sub-detection well, the purpose of detecting multiple factors separately in one main detection well can be achieved.

[0008] The position and distribution of the main detection holes can be consistent with the current standard 96-well plate, which is convenient for adding samples and cleaning using existing manual equipment or mechanical workbenches.

[0009] The location and distribution of the sub-detection wells can be consistent with the current standard 384-well plate, which is convenient for using a 384-well plate reader (including visible light, fluorescence or chemiluminescence plate reader) to record signals and process them using existing data processing software.

[0010] Furthermore, in the improved multi-well plate of the utility model for facilitating serum multi-factor mixed detection, the main detection well includes four sub-detection wells.

[0011] The main detection hole containing 4 sub-detection holes enables the operator to perform 4-factor detection at the same time, meeting the requirements for one-time 4-factor detection in small samples.

[0012] Furthermore, in the improved multi-well plate of the utility model for facilitating serum multi-factor mixed detection, the height of the transverse partition and the longitudinal partition are both between 1 / 10 and 1 / 3 of the depth of the main detection hole.

[0013] This arrangement ensures that the sub-detection holes can accommodate the antibodies to be detected, and at the same time facilitates the uniform flow and distribution of the sample liquid among the sub-detection holes.

[0014] Furthermore, in the improved multi-well plate of the utility model for facilitating serum multi-factor mixed detection, the depth of the sub-detection wells is 1 / 4 of the depth of the main detection wells.

[0015] The 1 / 4 depth sub-detection allows the liquid to flow and distribute evenly, while also preventing it from overflowing from the main detection hole.

[0016] Furthermore, in the improved multi-well plate of the utility model for facilitating serum multi-factor mixed detection, the main detection well includes 8 sub-detection wells.

[0017] The main detection hole containing 8 sub-detection holes is suitable for 8-factor detection. In addition, 8, 12 or even 16 factors can be detected simultaneously by combining two or more main detection plates.

[0018] The "combination method" is adopted to increase the number of detection factors. It only requires appropriate planning by changing the manual or mechanical workbench loading steps without further structural modification of the detection plate. In specific implementation, the sub-detection wells are 4 times the main detection wells. For example, the current standard 96-well plate has 384 sub-detection wells corresponding to the current standard 384-well positions after being improved.

[0019] Furthermore, in the improved multi-porous plate of the utility model for facilitating serum multi-factor mixed detection, the surface of the part where the transverse partition and the longitudinal partition are connected to each other is depressed downward to form a flow groove.

[0020] The provision of the flow slot enables the operator to conveniently inject the liquid and allows the liquid to flow between the sub-detection holes, thereby enabling the liquid to be quickly and evenly injected into the sub-detection holes.

[0021] Furthermore, in the improved multi-porous plate of the utility model which is convenient for serum multi-factor mixed detection, the transverse partition and the longitudinal partition are integrally formed with the plate body.

[0022] The transverse partition and the longitudinal partition formed integrally with the plate body not only facilitate the preparation of the improved detection hole plate, but also improve the strength of the detection hole plate.

[0023] Furthermore, the improved multi-well plate of the utility model that is convenient for serum multi-factor mixed detection, the plate body is a 384-well plate with 384 sub-detection wells.

[0024] This arrangement allows the operator to make slight improvements to the existing orifice plate mold to conveniently prepare the improved detection orifice plate.

[0025] Furthermore, the improved multi-well plate of the utility model which is convenient for serum multi-factor mixed detection has 24 main detection holes on the plate body, and each main detection hole has 16 sub-detection holes.

[0026] The above description is only an overview of the technical solution of the utility model. In order to more clearly understand the technical means of the utility model and implement it in accordance with the contents of the specification, the following embodiments of the utility model are described in detail. BRIEF DESCRIPTION OF THE DRAWINGS

[0027] Figure 1 This is a top view of the main detection hole.

[0028] Figure 2 yes Figure 1 Sectional view along AA direction.

[0029] Figure 3 It is a top view of the main detection hole with flow slot.

[0030] Figure 4 yes Figure 3 Cross-sectional view along the BB axis.

[0031] Figure 5 It is a detection well plate with a main detection well containing 4 sub-detection wells.

[0032] Figure 6 It is a detection well plate with a main detection well containing 8 sub-detection wells.

[0033] Figure 7 It is a detection well plate with a main detection well containing 16 sub-detection wells.

[0034] In the figure, there is a plate body 1, a main detection hole 2, a transverse partition 3, a longitudinal partition 4, a sub-detection hole 5, and a flow slot 6. DETAILED DESCRIPTION

[0035] The following is a further detailed description of the specific implementation of the present invention in conjunction with the accompanying drawings and examples. The following examples are used to illustrate the present invention, but are not intended to limit the scope of the present invention.

[0036] Example 1: See Figures 1 to 7 The improved multi-well plate for serum multi-factor mixed detection of this embodiment includes a plate body 1, and a plurality of main detection holes are provided on the surface of the plate body. 2. The main detection hole includes a plurality of transverse partitions 3 and longitudinal partitions 4, which are respectively connected to the inner wall of the main detection hole, and the height of the transverse partitions and longitudinal partitions is lower than the depth of the main detection hole. The transverse partitions and longitudinal partitions divide the main detection hole into a plurality of sub-detection holes 5 whose depth is less than that of the main detection hole.

[0037] The advantage of the improved multi-well plate for serum multi-factor mixed detection is that its main detection hole is divided into a plurality of sub-detection holes with a depth lower than that of the main detection hole by a relatively low transverse partition and a longitudinal partition. Thus, during multi-factor detection, the operator only needs to add the sample (including serum or culture fluid, mixed secondary antibody, tertiary antibody, color developer and cleaning fluid, etc.) once so that the height of the liquid is higher than the depth of the sub-detection hole, and the sample liquid can be easily and evenly injected into each sub-detection hole, which is very convenient to operate.

[0038] Among them, the main detection hole is similar to the detection hole of the conventional orifice plate, both of which are grooves with consistent depth and flush surface.

[0039] The sub-detection holes are formed by dividing the main detection hole with a plurality of transverse partitions and longitudinal partitions. The bottom surfaces of the transverse partitions and longitudinal partitions are both arranged on the bottom surface of the main detection hole, and the two ends thereof are connected to the inner wall of the main detection hole, or the surfaces of adjacent transverse partitions and longitudinal partitions. Since the height of the transverse partitions and longitudinal partitions is lower than the depth of the main detection hole, the depths of the plurality of sub-detection holes formed by the divisions are all lower than the depth of the main detection hole. At the same time, since a single main detection hole has a plurality of sub-detection holes, the aperture of the main detection hole is larger than that of the original orifice plate. During multi-factor detection, when injecting the sample liquid, the operator only needs to ensure that the surface of the liquid is higher than the surface of the sub-detection hole to achieve uniform injection into each sub-detection hole, which is more convenient to operate.

[0040] The height of the transverse and longitudinal partitions is between 1 / 10 and 1 / 3 of the depth of the main detection hole, as long as it can accommodate the antibody to be detected and facilitate the uniform flow and distribution of the sample liquid between the sub-detection holes. In this embodiment, the sub-detection hole is about 1 / 4 of the depth of the main detection hole. The main detection hole can accommodate 100 microliters of solution, while the sub-detection hole can accommodate 10 microliters of solution.

[0041] Multiple transverse and longitudinal partitions can be set in the main detection hole, and the number of sub-detection holes in a single main detection hole can be changed according to demand. However, the number, position and distribution of the sub-detection holes must match the existing sample pipette and plate reader.

[0042] For example, a transverse partition and a longitudinal partition may be vertically and cross-arranged in the main detection hole, and the transverse partition and the longitudinal partition may evenly divide the main detection hole into four sub-detection holes.

[0043] The design of sub-detection holes with other numbers of main detection holes is similar and will not be described in detail.

[0044] The above-mentioned transverse partition and longitudinal partition can be integrally formed with the plate body, and the size of the sub-detection hole can be consistent with the detection hole size of conventional detection well plates such as existing 96-well plates or 384-well plates, so that the operator can make slight improvements to the existing well plate mold to conveniently prepare the improved detection well plate.

[0045] In order to facilitate the injection of liquid and allow the liquid to flow between the sub-detection holes so as to be quickly and evenly injected into the sub-detection holes, the surface of the portion where the transverse partition and the longitudinal partition are connected to each other can be recessed downward to form a flow groove 6. The flow groove allows the liquid to flow conveniently between the sub-detection holes through the flow groove so as to keep the liquid level between the sub-detection holes consistent. During operation, the operator only needs to roughly align the portion where the flow groove is located to inject, which is relatively convenient.

[0046] The following uses the traditional Elisa method to detect inflammatory factors IL-1, IL-4, INF-γ, and TNF-α in mouse serum as an example to explain the specific method of using the improved detection plate for multi-factor detection. Among them, wells F1 to F4 are sub-detection wells in the same main detection well.

[0047] 1. Use a mechanical workbench or manual operation method matched with a traditional 384-well plate to add 10 microliters of 1-10ug / ml IL-1 antibody to F1 well, 10 microliters of 1-10ug / ml IL-4 antibody to F2 well, 10 microliters of 1-10ug / ml INF-γ antibody to F3 well, and 10 microliters of 1-10ug / ml TNF-α antibody to F4 well, and coat overnight at 4°C;

[0048] 2. After discarding the coated antibody, add 100ul of mouse serum sample containing four factors, IL-1, IL-4, INF-γ, and TNF-α, into the main detection well and incubate at 37°C for 1 hour;

[0049] 3. Discard the serum sample and wash with PBS washing solution 3-4 times;

[0050] 4. Add 100ul of mixed secondary antibodies containing 4 enzyme-labeled IL-1, IL-4, INF-γ, and TNF-α to the main detection well and incubate at 37°C for 1 hour;

[0051] 5. Discard the incubated secondary antibody and wash 3-4 times with PBS washing solution;

[0052] 6. Add precipitated color substrate (or corresponding precipitated fluorescent or chemiluminescent substrate) and allow color to develop for an appropriate time;

[0053] 7. Use a microplate reader (or chemiluminescence instrument) that can detect 384 wells to record the OD values ​​(or fluorescence or chemiluminescence values) of wells F1, F2, F3, and F4 and perform data analysis.

[0054] From the above description, it can be seen that the improved detection plate for multi-factor detection allows operators to use conventional detection equipment, such as conventional pipettes and ELISA readers, to achieve multi-factor detection. Compared with multi-factor detection through MSD, Luminex and other technical platforms, the detection cost (including special consumables and readers) and the professional requirements for operators are greatly reduced.

[0055] The above is only a preferred implementation of the utility model, which is used to assist those skilled in the art to implement the corresponding technical solution, and is not used to limit the scope of protection of the utility model. The scope of protection of the utility model is defined by the attached claims. It should be pointed out that for ordinary technicians in this field, on the basis of the technical solution of the utility model, several equivalent improvements and variations can be made, and these improvements and variations should also be regarded as the scope of protection of the utility model. At the same time, it should be understood that although this specification is described in accordance with the above-mentioned implementation mode, not every implementation mode contains only one independent technical solution. This narrative method of the specification is only for the sake of clarity. Those skilled in the art should regard the specification as a whole. The technical solutions of each embodiment can also be appropriately combined to form other implementation modes that can be understood by those skilled in the art.

Claims

1. An improved multi-well plate for facilitating serum multi-factor mixed detection, comprising a plate body (1), a plurality of main detection holes (2) on the surface of the plate body, characterized in that: The main detection hole includes a plurality of transverse partitions (3) and longitudinal partitions (4), which are respectively connected to the inner wall of the main detection hole. The height of the transverse partitions and the longitudinal partitions is lower than the depth of the main detection hole. The transverse partitions and the longitudinal partitions divide the main detection hole into a plurality of sub-detection holes (5) whose depths are less than the main detection hole.

2. The improved multi-well plate for facilitating serum multi-factor mixed detection according to claim 1, characterized in that: The main detection hole includes four sub-detection holes.

3. The improved multi-well plate for facilitating serum multi-factor mixed detection according to claim 1, characterized in that: The heights of the transverse partition and the longitudinal partition are both between 1 / 10 and 1 / 3 of the depth of the main detection hole.

4. The improved multi-well plate for facilitating serum multi-factor mixed detection according to claim 3, characterized in that: The depth of the sub-detection hole is 1 / 4 of the depth of the main detection hole.

5. The improved multi-well plate for facilitating serum multi-factor mixed detection according to claim 1, characterized in that: The main detection hole includes 8 sub-detection holes.

6. The improved multi-well plate for facilitating serum multi-factor mixed detection according to claim 1, characterized in that: The surface of the portion where the transverse partition and the longitudinal partition are connected to each other is concave downward to form a flow groove (6).

7. The improved multi-well plate for facilitating serum multi-factor mixed detection according to claim 1, characterized in that: The transverse partition and the longitudinal partition are integrally formed with the plate body.

8. The improved multi-well plate for facilitating serum multi-factor mixed detection according to claim 1, characterized in that: The plate body is a 384-well plate with 384 sub-detection wells.

9. The improved multi-well plate for facilitating serum multi-factor mixed detection according to claim 8, characterized in that: The plate body is provided with 24 main detection holes, and each main detection hole is provided with 16 sub-detection holes.

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