Periodic acid-Schiff (pas) staining solution (chemical staining method)

A chemical dyeing and periodic acid technology, applied in the biological field, can solve the problems of unsatisfactory dyeing effect, low sensitivity, time-consuming detection, etc., and achieve the effects of uniform and bright color, high detection sensitivity, and fast fixation time.

Active Publication Date: 2018-05-22
SHANGHAI SUNBIO TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The Schiff reagent in the commercially available PAS staining kit is mainly prepared by the above method. When used for PAS staining, the sensitivity is not high and the staining effect is not ideal for cells with low sugar content, and it is often necessary to develop color with glycogen in blood cells. The reaction is more than 1 hour, and the detection is quite time-consuming
In addition, existing PAS methods or commercially available PAS kits also have the defect of high false negatives

Method used

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  • Periodic acid-Schiff (pas) staining solution (chemical staining method)
  • Periodic acid-Schiff (pas) staining solution (chemical staining method)
  • Periodic acid-Schiff (pas) staining solution (chemical staining method)

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Effect test

Embodiment 1

[0033] Embodiment 1: Composition and preparation method of the kit of the present invention

[0034] PAS fixative: 0.25% potassium permanganate: methanol: 40% formaldehyde = 1:4:1 (v / v / v);

[0035] PAS Ⅰ solution: 1% periodic acid solution;

[0036]PAS II solution: Weigh 1g of basic fuchsin and add it to boiling 200mL distilled water, stir until the fuchsin is completely dissolved, cool the solution to 50°C and filter. Add 20ml of 1mol / L acetic acid and cool overnight. Use acetic acid to adjust the pH of the solution to 2.00±0.20, stir for 1h, continue to drop acetic acid until the pH is stable to 2.00±0.20, add 1-1.5g of sodium metabisulfite to the above solution, stir to dissolve, stand in the dark for 24h, add 1.0 - Stir 1.5g of activated carbon for 2 minutes, filter, and store in a refrigerator;

[0037] PAS counterstain: Mayer's hematoxylin stain.

Embodiment 2

[0038] Embodiment 2: composition and preparation method of the kit of the present invention

[0039] PAS fixative: 0.25% potassium permanganate: methanol: 40% formaldehyde = 1:4:1 (v / v / v);

[0040] PAS Ⅰ solution: 1% periodic acid solution;

[0041] PAS II solution: Weigh 1g of basic fuchsin and add it to boiling 200mL distilled water, stir until the fuchsin is completely dissolved, cool the solution to 50°C and filter. Add 20ml of 1mol / L formic acid and cool overnight. Use formic acid to adjust the pH of the solution to 2.00±0.20, stir for 1h, continue to drop formic acid until the pH is stable to 2.00±0.20, add 1-1.5g of sodium metabisulfite to the above solution, stir to dissolve, let stand in the dark for 24h, add 1.0 - Stir 1.5g of activated carbon for 2 minutes, filter, and store in a refrigerator;

[0042] PAS counterstain: Mayer's hematoxylin stain.

Embodiment 3

[0043] Embodiment 3: composition and preparation method of the kit of the present invention

[0044] PAS fixative: 0.25% potassium permanganate: methanol: 40% formaldehyde = 1:4:1 (v / v / v);

[0045] PAS Ⅰ solution: 1% periodic acid solution;

[0046] PAS II solution: Weigh 1g of basic fuchsin and add it to boiling 200mL distilled water, stir until the fuchsin is completely dissolved, cool the solution to 50°C and filter. Add 20ml of 0.5mol / L oxalic acid and cool overnight. Use oxalic acid to adjust the pH of the solution to 2.00±0.20, stir for 1 hour, continue to drop oxalic acid until the pH is stable to 2.00±0.20, add 1-1.5g sodium metabisulfite to the above solution, stir to dissolve, and let it stand in the dark After 24 hours, add 1.0-1.5g of activated carbon, stir for 2 minutes, filter, and store in a refrigerator;

[0047] PAS counterstain: Mayer's hematoxylin stain.

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Abstract

The invention relates to the field of biotechnology, and discloses a periodic acid-Schiff chemical staining kit. The kit of the present invention includes PAS fixative solution, PAS I solution, PAS II solution and PAS counterstaining solution; wherein, the PAS fixative solution is composed of potassium permanganate, methanol and formaldehyde; the PAS I solution is iodine Acid solution; the PAS II solution is Schiff reagent. The invention adjusts the composition of the fixative, thereby shortening the sample fixing time and improving the sample fixing and dyeing effect. On this basis, the present invention further improves the preparation mode of the Schiff reagent, achieves the effect of improving sensitivity, and further shortens the entire PAS detection time. At the same time, the detection result of the kit provided by the invention is more accurate, effectively reducing the occurrence of false negative rate results.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a periodic acid-Schiff (PAS) staining solution (chemical staining method), namely a periodic acid-Schiff chemical staining kit. Background technique [0002] Periodic acid-Schiff (PAS) staining is one of the commonly used chemical staining methods for blood cells. It plays an important role in the diagnosis, differential diagnosis and curative effect observation of certain blood diseases such as red blood cell line, white blood cell line, and lymphatic system. The staining principle is: sugar substances containing ethylene glycol in cells can be oxidized by periodic acid to form dialdehyde substances, which combine with colorless fuchsin in Schiff reagent to form purple-red precipitates. The color depth is proportional to the content of ethylene glycol. With the development of medical experiment technology, the scope of application of PAS is more extensive, such as to prove and iden...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N1/30
Inventor 谢永华朱美萍许付
Owner SHANGHAI SUNBIO TECH
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