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Buffer solution for immunohistochemical detection quality control product as well as preparation method and application of buffer solution

A technology of immunohistochemistry and buffer solution, which is applied in application, measurement device, preservation of human or animal body, etc. It can solve the problems of tissue/cell inhomogeneity, cross-contamination, easy stacking, etc., and achieve ultra-long-term preservation and volatilization Rapid, non-diffusing effect

Active Publication Date: 2022-06-28
HANGZHOU BIOLYNX TECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] In order to solve at least one of the above-mentioned technical problems, the present invention provides a new type of buffer, which has the characteristics of rapid volatilization, difficult diffusion, stable properties, and ultra-low temperature storage resistance by adjusting the buffer solvent, viscosity, adding stabilizers, and antibacterial agents , which solves the problems of cross-contamination and uneven tissue / cell stacking in the existing technology; in addition, by changing the formula of the matching solvent, the buffer solution can be used to prepare tissue / cell quality control products, which can ensure longer-term stability in the buffer solution. Stable tissue / cell antigens for ultra-long-term storage

Method used

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  • Buffer solution for immunohistochemical detection quality control product as well as preparation method and application of buffer solution
  • Buffer solution for immunohistochemical detection quality control product as well as preparation method and application of buffer solution
  • Buffer solution for immunohistochemical detection quality control product as well as preparation method and application of buffer solution

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0062] Embodiment 1 Dissolution scheme optimization of carbomer

[0063] In order to dissolve carbomer, the inventors designed the following schemes:

[0064] Option One:

[0065] Weigh 40g of absolute ethanol and about 50g of deionized water, stir for 30sec and mix well, weigh 1g of Carbomer 940 and slowly disperse it into the ethanol and deionized water solution. Stir or shake at 120RPM on a horizontal shaker for 30min until the carbomer is fully dissolved, and it is a transparent slightly viscous liquid with bubbles. The measured pH value is 6.1. About 0.2 g of triethanolamine was added dropwise, and the pH value was adjusted to 7.2. At this time, a transparent viscous liquid with bubbles appeared. Add 10 mg of sodium azide and make up the balance with deionized water, continue to place on a horizontal shaker at 120 RPM at room temperature for overnight defoaming to form a clear and stable gel-like buffer.

[0066] Option II:

[0067] Weigh 1g of Carbomer 940 and slowly...

Embodiment 2

[0072] Example 2 Buffer for immunohistochemical detection quality control product and its preparation

[0073] To verify the performance of different buffers, prepare different buffers according to Table 1.

[0074] The content (g) and pH value of each component in the buffer solution of Table 1

[0075] buffer R1 R2 A1 A2 A3 A4 B1 B2 B3 B4 Carbomer 940 / / 0.2 / 1.0 / 0.2 / 1.0 / Carbomer 980 / / / 0.2 / 0.5 / 0.2 / 0.5 anhydrous ethanol / 35 35 35 40 40 / / / / isopropyl alcohol / / / / / / 30 30 35 35 Deionized water 100 65 margin margin margin margin margin margin margin margin Triethanolamine / / 0.1-0.2 0.1-0.2 0.1-0.2 0.1-0.2 0.1-0.2 0.1-0.2 0.1-0.2 0.1-0.2 Sodium azide / / 0.01 0.01 0.01 0.01 0.01 0.01 0.01 0.01 pH / / 7.2 7.2 7.2 7.2 7.2 7.2 7.2 7.2

[0076] The preparation methods of the above buffers are as follows: ...

Embodiment 3

[0084] Example 3 Distribution of cells in cell suspension prepared with different buffers

[0085] 1. Preparation of Cell Suspension

[0086] In each buffer prepared in Example 2, according to 10M (10×10 per mL of buffer) 6 The concentration of each) cells was added to the cells respectively, and different cell suspensions were prepared.

[0087] 2. Observation of Cell Distribution in Cell Suspension

[0088] For each cell suspension, 1 μL was dropped onto a glass slide and stained with Ki-67 antibody as the primary antibody for detection. It can be observed that the cell suspension prepared with buffer B2 does not spread after being added dropwise to the glass slide, and it only takes about 3 minutes to completely evaporate at room temperature. It is observed under a microscope that a uniform cell sheet can be formed ( figure 1 ), and the cell suspension quality control products prepared with buffers A1-A4 and B1-B4 performed basically the same; the cell suspension prepare...

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Abstract

The invention discloses a buffer solution for an immunohistochemical detection quality control product, and belongs to the technical field of immunohistochemical detection. The buffer solution is prepared from the following components in percentage by mass: 0.2 to 1 percent of carbomer, 30 to 40 percent of alcohol, 0.1 to 0.25 percent of triethanolamine, 0.005 to 0.015 percent of sodium azide and the balance of deionized water. By adjusting the solvent and viscosity of the buffer solution and adding the stabilizer and the antibacterial agent, the gel has the characteristics of rapid volatilization, difficult diffusion, stable property, ultralow temperature storage resistance and the like, and solves the problems of easy cross contamination, uneven tissues / cells and easy stacking in the prior art. Meanwhile, the buffer solution disclosed by the invention can ensure the stability of tissue / cell antigens in the buffer solution for a longer time, so that ultra-long-term preservation is realized. The preparation method of the buffer solution is simple and easy to operate, and the buffer solution is wide in application, can be suitable for different tissues or cells, and has huge popularization value.

Description

technical field [0001] The invention belongs to the technical field of immunohistochemistry, and particularly relates to a buffer for immunohistochemistry detection quality control substance and its preparation and application. Background technique [0002] Immunohistochemical detection, abbreviated as immunohistochemical detection, is a conventional immunological detection method that combines antigen-antibody specificity and develops color by chromogenic reagent. This method is mainly applied to the pathological detection of biological tissue slice samples. In the detection, a specific primary antibody is used to specifically bind to the antigen in the biological tissue section sample, and a general secondary antibody is further used to bind to the aforementioned primary antibody, and color is developed by a chromogenic molecule linked to the secondary antibody. The pathological state of the biological tissue section can be judged by analyzing the detected staining situat...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A01N1/02G01N33/531
CPCA01N1/0226G01N33/531
Inventor 胡旻李明振潘丽蔡宁
Owner HANGZHOU BIOLYNX TECH CO LTD
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