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Alpha-naphthol acetate esterase staining kit

A technology of naphthol acetate and naphthol acetate, which is applied in the field of α-naphthol acetate staining kits, can solve the problem of the large influence of NaF inhibition rate, the trouble of leukemia typing and identification, and the difficulty of directly distinguishing single nuclei Cells and promyelocytes, etc., to shorten the detection time, eliminate interference effects, and improve detection efficiency

Active Publication Date: 2016-06-08
SHANGHAI SUNBIO TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Existing staining methods can observe the activity of α-NAE, but it is difficult to directly distinguish monocytes and promyelocytes by existing staining methods when typing acute leukemia, and it is necessary to distinguish monocytes and granulocytes through NaF inhibition test. Cell line leukemia, and the judgment of NaF inhibition rate is greatly affected by human subjective factors, which often brings some troubles to leukemia typing and identification

Method used

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  • Alpha-naphthol acetate esterase staining kit
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  • Alpha-naphthol acetate esterase staining kit

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0034] Embodiment 1: kit of the present invention and dyeing method

[0035] 1. Composition and preparation method of the kit of the present invention

[0036] Fixative: Weigh 20mgNa 2 HPO 4 12H 2 O, 100mgKH 2 PO 4 , add 30ml of distilled water, add 45ml of acetone and 25ml of formaldehyde after dissolving, adjust the pH value to 6.6 after mixing.

[0037] α-naphthyl acetate solution: Weigh 100 mg of α-naphthyl acetate and dissolve it in 10 ml of 50% acetone aqueous solution.

[0038] Azo dye solution: weigh 300mg of basic blue 54 and dissolve in 10ml of distilled water, the color is dark red.

[0039] Buffer (base solution): 0.067M phosphate buffer at pH 7.6.

[0040] Preparation method: Liquid A: Weigh 2.388gNa 2 HPO 4 12H 2 Add distilled water to dissolve O to 100ml; B solution: weigh 0.908gKH 2 PO 4 Add distilled water to dissolve to 100ml. Take 87ml of liquid A and 13ml of liquid B and mix well to get the product.

[0041] Rinsing solution: 0.1M acetic acid-...

Embodiment 2

[0047] Example 2: Existing conventional α-NAE staining method

[0048] 1. Composition and preparation method of conventional staining reagents

[0049] Action solution: 50ml of 0.05M phosphate buffer (pH7.6), 100mg of α-naphthol acetate dissolved in 1ml of 50% acetone aqueous solution, shake fully until most of the initial turbidity disappears, add diazonium salt (solid blue B) Shake 50mg, filter and use.

[0050] Counterstain solution: 10g / L methyl green aqueous solution.

[0051] 2. Dyeing method

[0052] (1) Fixation: Fresh smears were fumigated with formaldehyde for 5 to 10 minutes.

[0053] (2) Rinse with water for 5 minutes.

[0054] (3) Put into the working solution (37°C) for 1 hour, and wash with water.

[0055] (4) Counterstain with 10g / L methyl green aqueous solution for 5 minutes.

[0056] (5) Microscopic examination after washing and drying.

Embodiment 3

[0057] Embodiment 3: the comparison of kit staining of the present invention and existing routine staining method

[0058] Acute monocytic leukemia (M5) and acute promyelocytic leukemia (M3) patient blood cells were stained by using the kit of Example 1 of the present invention and according to the existing conventional staining method of Example 2, the results are shown in Figure 1-Figure 4 .

[0059] The dyeing result of the present invention shows that in the M5 type leukemia blood cells, the nucleus of the mononuclear cell line is bright reddish purple, and the cytoplasm is reddish purple, and α-naphthol acetate esterase in the mononuclear cells forms black granule precipitation; The nuclei of granulocytic cells are brown or dark brown, and black granular precipitates are formed at the α-naphthol acetate esterase in the cytoplasm.

[0060] Existing method dyeing result, in the M5 type leukemia blood cell, the nucleus of the monocyte cell line is green, and the α-naphthol...

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Abstract

The invention relates to the technical field of biology, and discloses an alpha-naphthol acetate esterase staining kit. The kit comprises a fixing liquid, an alpha-naphthol acetate solution, an azo dye solution, a buffer liquid, and a rinsing liquid; wherein the azo dye solution is an alkali blue 54 water solution. The azo dyes such as fast blue B salt, fast violet salt, and the like in the conventional alpha-NAE staining method are replaced by the alkali blue 54; the staining time is shorten to about 20 minutes; moreover, after the staining, the mononuclear cells, immature granular cells, mature granular cells, lymphocyte and erythrocyte can be differentiated in the microscopic examination area, the re-staining of cell nucleus is not needed, furthermore, monocytic leukemia and granulocytic leukemia can be diagnosed quickly without the assistance of NaF inhibition tests, and the diagnosis method using the kit is faster and more convenient, compared with the conventional method.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to an α-naphthyl acetate esterase (α-NAE) staining liquid (chemical staining method), namely an α-naphthyl acetate esterase staining kit. Background technique [0002] Cytochemical staining technology is a combination of cytology and chemistry, based on morphology, through chemical staining, to perform qualitative, quantitative and localized analysis of enzymes, lipids, sugars, proteins, nucleic acids and other components in cells under a microscope of a technique. This technology is applied to blood and bone marrow smears and lymph node tissue sections, and plays an important role in the identification of blood cells, diagnosis and differential diagnosis of blood diseases. At present, commonly used cytochemical stains related to blood system diseases include peroxidase staining, glycogen staining, esterase staining, alkaline phosphatase staining, acid phosphatase staining, and iron st...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/44
CPCC12Q1/44C12Q2334/30
Inventor 谢永华朱美萍许付
Owner SHANGHAI SUNBIO TECH
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