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Improved methenamine silver-masson staining method

A silver-massone and silver-staining technology, which is applied in the field of hexaamine silver-massone overstaining, can solve problems such as high background color, difficult control of time and temperature, and easy overstaining of tissues

Inactive Publication Date: 2014-07-16
NANFANG HOSPITAL OF SOUTHERN MEDICAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] At present, professionals at home and abroad have tried many ways to improve the commonly used basement membrane silver staining methods, including Gomori method, Jones method, Yajima (Jenos) improved method, Gordon-Sweet method, Janes method, etc., in an attempt to obtain better results. The quality of basement membrane staining, but there are still the following problems: the tissue is prone to overstaining in silver staining; time and temperature are difficult to control; the background color is too high;

Method used

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Examples

Experimental program
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Effect test

Embodiment 1

[0046] Embodiment 1: hexamine silver masson overdyeing method of the present invention

[0047] 1. Reagent formula:

[0048] 1.1 Silver dyeing formula:

[0049] Silver stain A liquid: distilled water 230ml, 5% borax 45ml, 4% boric acid 4ml, 15% hexamethylenetetramine 60ml. The liquid is stored in a refrigerator at 4°C and is valid for 2 months. Reconstitution is required after 2 months.

[0050] Silver stain B solution: 100ml distilled water, 10g silver nitrate. The liquid needs to be stored in a refrigerator at 4°C away from light. If black precipitation is found in the liquid, it needs to be reconstituted.

[0051] 1.2.15% borax: 100ml distilled water, 5g borax. Store the liquid at room temperature, away from light.

[0052] 1.2.2 15% hexamethylenetetramine: 100ml of distilled water, 15g of hexamethylenetetramine, the liquid is stored in a refrigerator at 4°C, and it needs to be reconstituted if precipitation occurs.

[0053] 1.2.34% boric acid: 100ml distilled water,...

Embodiment 2

[0078] Embodiment 2: the comparison of the hexamine silver masson overstaining method of the present invention and the kidney tissue hexamine silver staining of major hospitals and medical inspection institutions at home and abroad

[0079] figure 1 It is a kidney tissue section staining diagram of the improved hexaaminesilvermasone overstaining method described in the present invention. Such as figure 1 The improved Hexamine Silver Masson overstaining method can clearly outline the black lines of the glomerular basement membrane, renal tubular basement membrane, and vascular basement membrane in the kidney tissue, and the immune complexes outside the basement membrane and renal tubules The cells are brightly stained, the stained background is clean, and the colors are vivid.

[0080] From the perspective of staining effect, this staining method is better than Jones-HE silver staining method ( figure 2 ), Gomori-masson silver staining ( image 3 ), Yajima (Jenos) modified...

Embodiment 3

[0081] Embodiment 3: the amine silver reaction comparison of different experimental water under the same conditions

[0082] Choose single-distilled water, double-distilled water, the first-grade water in the analysis laboratory treated by the water treatment system, the double-distilled treated water after the first-grade water in the analysis laboratory, and 30 samples of electronic-grade ultrapure water, and add the water of the present invention respectively. For silver staining solution A and silver staining solution B, the amine-silver reaction was carried out in an oven at 70°C for 1.5 hours. Each sample was placed in a 752-type ultraviolet spectrophotometer for measurement, and the wavelength was set at 680nm. Take the average value of 30 cases of measurement results, adjust to 0 with electronic grade ultra-pure mercury staining reagent solution, and the comparison results are as follows after statistical analysis Figure 6 As shown, it can be seen that the use of ele...

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Abstract

The invention relates to an improved methenamine silver-masson staining method. The method comprises the following steps of preparing a silver staining reagent; fixing tissue slices by Bouin liquor; immersing the slices into 2.5% iodine, 5% sodium thiosulfate and 1.5% periodic acid sequentially; carrying out dip dyeing on the slices by silver staining liquor A and silver staining liquor B till that the clear black is displayed at a basement membrane; soaking the slices by 3% sodium thiosulfate; carrying out dip dyeing by hematoxylin; differentiating by hydrochloric acid alcohol, and washing back to blue; carrying out dip dyeing by Ponceaux dye liquor; soaking the slices by 1% phosphomolybdic acid; and dewatering by gradient alcohol, putting into xylene to be transparent, and mounting by neutral gum. The invention provides the methenamine silver-masson staining method which is reasonable in reagent preparation, good in dyeing effect, short in time, stable in performance and bright-colored in resistdye color and luster, the effect of the method which is better than that of the prior art can be obtained on trial, and the problems that the tissue is easy to overstrain, the time and the temperature are hard to control, the background color is over high, and the other resistdye is not bright in dyeing color and the like in the silver staining can be changed.

Description

technical field [0001] The invention relates to a hexaminesilvermason overstaining method, which is one of the necessary special staining methods for pathological diagnosis of kidney tissue puncture biopsy. In particular, the present invention relates to an improved overstaining method of hexaamine argentasone, which can be used to display changes such as glomeruli, tubules, vascular basement membranes, tissue immune complex deposition and fibrosis in kidney tissue. Background technique [0002] The hexamine silver-masson overstaining method is a staining method for kidney tissue sections evolved from the hexamine silver method of mold staining. It is a superimposed staining of the two staining methods of hexamine silver and masson. In glomerular disease, often results in changes in the glomerular basement membrane, a highly aqueous gelatinous lining composed mainly of mucopolysaccharides and proteins that is permeable and often associated with fine connective tissue The fi...

Claims

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Application Information

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IPC IPC(8): G01N1/30
Inventor 周展眉王国保曹维杨满球
Owner NANFANG HOSPITAL OF SOUTHERN MEDICAL UNIV
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