Concentrated buffer cleaning solution

Abstract

The invention discloses a concentrated buffer cleaning solution which mainly comprises the following raw materials: disodium hydrogen phosphate, sodium dihydrogen phosphate, Tween-20, propylene glycol, sodium chloride, trihydroxymethyl aminomethane (Tris) and pure water, and the pH value range of the concentrated buffer cleaning solution is 8.9-9.1. According to the invention, a unique lysis and buffer combination system is adopted, and various types of histological and cytological samples and the like can be compatible. A high (pH value) antigen buffer cleaning solution Buffer can be compatible with various histological and cytological sample types including fresh tissues or frozen cells, hydrothorax and ascites, tissue exudate and the like, and free nucleic acid and nucleic acid binding protein in various histological and cytological samples are rapidly and thoroughly separated under the synergistic effect of protease K; then, the separated free nucleic acid is efficiently combined with the antibody under the high-salt condition provided by the adsorption combination buffer solution, and denatured protein, lipid and other impurities are prevented from blocking a probe of a machine.

Description

Technical Field

[0001] The invention relates to the technical field of pathological diagnosis, in particular to a concentrated buffer cleaning solution. Background Art

[0002] The detection technology of immunohistochemical staining in pathological diagnosis has developed rapidly in pathological diagnosis. It is the gold standard for immunohistochemical staining detection in the pathology department. Immunohistochemical staining is an important pathological auxiliary diagnosis method for clinical surgical medication. In the application of clinical guidance of medication, it has received increasing attention. Immunohistochemical staining technology has developed rapidly in recent years, and with the application and promotion of fully automatic immunohistochemical staining machines, the requirements for staining quality have also increased. Among them, concentrated buffer washing fluid plays an extremely important role in the entire immunohistochemical staining process. The quality of tissue washing conditions directly determines the quality of the final staining results. Due to its own structural characteristics, the container used to hold the concentrated buffer washing fluid of the fully automatic staining instrument is small in volume, and the tissue section cannot be completely immersed in the antigen buffer washing fluid like manual washing. In addition, the fully automatic immunohistochemical staining instrument needs to be washed at a higher temperature (about 100°C) when using the antigen buffer washing fluid. Therefore, the antigen buffer washing fluid is easy to evaporate, which further reduces the volume of the antigen buffer washing fluid, making its tissue washing effect relatively poor, and also seriously affecting the effect of subsequent immunohistochemical staining.

[0003] Therefore, we propose a concentrated buffer cleaning solution to solve the above problems. Summary of the invention

[0004] The object of the present invention is to provide a concentrated buffer cleaning solution to solve the problem that the volume of the current antigen buffer cleaning solution is reduced, resulting in relatively poor tissue cleaning effect and also seriously affecting the effect of subsequent immunohistochemical staining.

[0005] To achieve the above object, the present invention provides the following technical solutions:

[0006] The invention discloses a concentrated buffer cleaning solution, the raw materials of which mainly include: disodium hydrogen phosphate, sodium dihydrogen phosphate, Tween-20, propylene glycol, sodium chloride, tris(hydroxymethyl)aminomethane (Tris) and pure water, and the pH value of the concentrated buffer cleaning solution ranges from pH 8.9 to pH 9.1.

[0007] In a further embodiment, each 1000 mL of the concentrated buffer cleaning solution comprises: 2.2-3.25 g of disodium hydrogen phosphate, 0.2-0.5 g of sodium dihydrogen phosphate, 5-36 mL of Tween-20, 5-50 mL of propylene glycol, 0.3-0.5 mL of tris (hydroxymethyl) aminomethane (Tris), 2.5-7.5 g of sodium chloride and the remainder of purified water. The pH value of the high (pH value) antigen buffer cleaning solution Buffer ranges from pH 8.9 to 9.0.

[0008] In a further embodiment, each 1000 mL of the concentrated buffer cleaning solution comprises: 2.2-3.25 g of disodium hydrogen phosphate, 0.2-0.5 g of sodium dihydrogen phosphate, 5-36 mL of Tween-20, 5-50 mL of propylene glycol, 0.3-0.5 mL of tris (hydroxymethyl)aminomethane (Tris), 2.5-7.5 g of sodium chloride and the remainder of purified water. The pH value of the concentrated buffer cleaning solution ranges from pH 9 to 9.2.

[0009] Compared with the prior art, the present invention has the following beneficial effects:

[0010] The unique lysis and binding buffer system is compatible with various types of histological and cytological samples. The concentrated buffer washing solution is compatible with various types of histological and cytological samples, including fresh tissue or frozen cells, pleural effusion, tissue exudate, etc. Under the synergistic effect of proteinase K, the free nucleic acids and nucleic acid binding proteins in various histological and cytological samples are quickly and thoroughly separated; then the high salt conditions provided by the adsorption binding buffer allow the separated free nucleic acids to efficiently bind to antibodies and prevent impurities such as denatured proteins and lipids from clogging the probe of the machine; finally, a multi-step washing operation is used to remove protein, pigment, lipid and other inhibitory impurities to the maximum extent, and it is used in conjunction with the fully automatic immunohistochemical staining machine. The negative pressure device can increase the sample processing volume by 20ml and can process dozens of samples at the same time. It is easy to operate and only takes 10 minutes to 1.5 hours to complete a batch cleaning, reducing clinical costs. The concentrated buffer cleaning solution can be used in a large number of tissue section samples to clean free nucleic acids in tissue sections. The use of high-efficiency antigen-antibody binding target fragments can effectively improve the expression rate of antigens. The volume of the cleaning solution Buffer can be flexibly changed between 150μL and 200μL. The high (pH value) antigen buffer cleaning solution Buffer after 20 times dilution can meet the needs of immunohistochemical staining quality in multi-field research and applications of molecular biology, and has broad application prospects and market promotion value. DETAILED DESCRIPTION

[0011] The technical solutions in the embodiments of the present invention are described clearly and completely below. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of them. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0012] The raw materials of a concentrated buffer cleaning solution mainly include disodium hydrogen phosphate, sodium dihydrogen phosphate, Tween-20, propylene glycol, sodium chloride, tris(hydroxymethyl)aminomethane (Tris) and pure water, and the pH value of the concentrated buffer cleaning solution ranges from pH 8.9 to pH 9.1.

[0013] Example 1

[0014] Each 1000 mL of the concentrated buffer cleaning solution comprises: 2.2-3.25 g of disodium hydrogen phosphate, 0.2-0.5 g of sodium dihydrogen phosphate, 5-36 mL of Tween-20, 5-50 mL of propylene glycol, 0.3-0.5 mL of tris(hydroxymethyl)aminomethane (Tris), 2.5-7.5 g of sodium chloride and the remainder of purified water. The pH value of the concentrated buffer cleaning solution ranges from pH 8.9 to 9.0.

[0015] Example 2

[0016] Each 1000 mL of the concentrated buffer cleaning solution comprises: 2.2-3.25 g of disodium hydrogen phosphate, 0.2-0.5 g of sodium dihydrogen phosphate, 5-36 mL of Tween-20, 5-50 mL of propylene glycol, 0.3-0.5 mL of tris(hydroxymethyl)aminomethane (Tris), 2.5-7.5 g of sodium chloride and the remainder of purified water. The pH value of the concentrated buffer cleaning solution ranges from pH 9 to 9.2.

[0017] Concentration comparison table of specific buffers:

[0018] 1. Tris-HCl buffer (0.05 M, 25°C)

[0019] Mix 50 ml of 0.1 M Tris solution and X ml of 0.1 N hydrochloric acid, and dilute to 100 ml with water.

[0020]

[0021]

[0022] Tris(hydroxymethyl)aminomethane(Tris)HOCH2 CH2OH HOCH2 NH2

[0023] Molecular weight = 121.14;

[0024] 0.1M solution is 12.114 g / L. Tris solution can absorb carbon dioxide from the air

[0025] 2. Phosphate buffer

[0026] (1) Disodium hydrogen phosphate-sodium dihydrogen phosphate buffer (0.2)

[0027]

[0028]

[0029] Na2HPO4·2H2O molecular weight = 178.05, 0.2mol / L solution is 85.61 g / L. Na2HPO4·2H2O molecular weight = 358.22, 0.2mol / L solution is 71.64 g / L. Na2HPO4·2H2O molecular weight = 156.03, 0.2mol / L solution is 31.21 g / L. (2) Disodium hydrogen phosphate-potassium dihydrogen phosphate buffer (1 / 15mol / L)

[0030]

[0031] 3. Sodium carbonate-sodium bicarbonate buffer (0.1M)

[0032] Ca 2+ Mg 2+ Do not use when present

[0033]

[0034]

[0035] Na2CO2·10H2O molecular weight = 286.2; 0.1M solution is 28.62 g / L.

[0036] N2HCO3 molecular weight = 84.0; 0.1M solution is 8.40 g / L.

[0037] 4. "PBS" buffer

[0038]

[0039] It will be apparent to those skilled in the art that the present invention is not limited to the details of the exemplary embodiments described above and that the present invention can be implemented in other specific forms without departing from the spirit or essential features of the present invention. Therefore, the embodiments should be considered exemplary and non-restrictive in all respects, and the scope of the present invention is defined by the appended claims rather than the above description, and it is intended that all changes falling within the meaning and scope of the equivalent elements of the claims be included in the present invention.

[0040] In addition, it should be understood that although the present specification is described according to implementation modes, not every implementation mode contains only one independent technical solution. This description 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 in each embodiment may also be appropriately combined to form other implementation modes that can be understood by those skilled in the art.

Claims

1. A concentrated buffer cleaning solution, characterized in that: The raw materials mainly include disodium hydrogen phosphate, sodium dihydrogen phosphate, Tween-20, propylene glycol, sodium chloride, tris(hydroxymethyl)aminomethane (Tris) and pure water, and the pH value range of the concentrated buffer cleaning solution is pH 8.9-9.

1.

2. The concentrated buffer cleaning solution according to claim 1, characterized in that: Each 1000mL of the concentrated buffer cleaning solution comprises: 2.2-3.25g of disodium hydrogen phosphate, 0.2-0.5g of sodium dihydrogen phosphate, 5-36mL of Tween-20, 5-50mL of propylene glycol, 0.3-0.5mL of tris(hydroxymethyl)aminomethane (Tris), 2.5-7.5g of sodium chloride and the remainder of purified water. The pH value of the high (pH value) antigen buffer cleaning solution Buffer ranges from pH 8.9 to 9.

0.

3. The concentrated buffer cleaning solution according to claim 1, characterized in that: Each 1000 mL of the concentrated buffer cleaning solution comprises: 2.2-3.25 g of disodium hydrogen phosphate, 0.2-0.5 g of sodium dihydrogen phosphate, 5-36 mL of Tween-20, 5-50 mL of propylene glycol, 0.3-0.5 mL of tris(hydroxymethyl)aminomethane (Tris), 2.5-7.5 g of sodium chloride and the remainder of purified water. The pH value of the concentrated buffer cleaning solution ranges from pH 9 to 9.2.

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