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Dyeing method for tissue glycogen

A dyeing method and technology of glycogen, applied in the biological field, can solve the problem of whether glycogen accumulates and where it accumulates, which cannot be easily and effectively preserved

Inactive Publication Date: 2017-12-19
GUANGZHOU KINGMED DIAGNOSTICS CENT
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] Based on this, it is necessary to provide a tissue glycogen staining method for the problems that the existing technology cannot simply and effectively preserve glycogen, judge whether glycogen is accumulated and where it accumulates

Method used

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  • Dyeing method for tissue glycogen
  • Dyeing method for tissue glycogen
  • Dyeing method for tissue glycogen

Examples

Experimental program
Comparison scheme
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Embodiment 1

[0034] This embodiment provides a method for staining tissue glycogen, comprising the following steps:

[0035] (1) Section preparation: fix the freshly obtained target tissue with 2.5% (mass percent) neutral glutaraldehyde and 1.2% (mass percent) osmic acid, and then undergo routine procedures such as rinsing, dehydration, infiltration, resin embedding, and sectioning. The corresponding semi-thin slices were obtained through the transmission electron microscope section preparation process.

[0036] (2) Periodic acid oxidation: put the above-mentioned semi-thin slices on a 70°C roaster for 10 minutes, then add periodate solution (0.5-1.5% by mass) in a 38°C oven and place it for 13-20min (high Periodic acid can oxidize the polysaccharide ethylene glycol group in the cell into dialdehyde, and the periodate solution needs to be preheated, the preheating temperature is 38°C and the time is 12min); the temperature limited in this step can speed up the subsequent dyeing solution an...

Embodiment 2

[0043] This embodiment provides a method for staining tissue glycogen, comprising the following steps:

[0044] (1) Slice preparation: fix the freshly obtained target tissue with 2% neutral glutaraldehyde and 1.5% osmic acid, and then undergo conventional TEM slice preparation procedures such as rinsing, dehydration, infiltration, resin embedding, and sectioning to obtain Corresponding semi-thin slices.

[0045](2) Periodic acid oxidation: put the above semi-thin slices on a 70°C oven for 5 minutes, and then add periodic acid solution in a 35°C oven for 13 minutes (periodic acid can make the polysaccharide ethylene glycol in the cells base is oxidized to dialdehyde, and the periodate solution needs to be preheated. The preheating temperature is 35°C and the time is 10min); within 2 weeks to achieve the desired staining effect; periodate solution is a solution that is prepared within 2 weeks and stored in the refrigerator. When adding periodate solution, attention should be pa...

Embodiment 3

[0052] This embodiment provides a method for staining tissue glycogen, comprising the following steps:

[0053] (1) Slice preparation: fix the freshly obtained target tissue with 3% neutral glutaraldehyde and 1.0% osmic acid, and then undergo conventional TEM slice preparation procedures such as rinsing, dehydration, infiltration, resin embedding, and sectioning to obtain Corresponding semi-thin slices.

[0054] (2) Periodic acid oxidation: put the above semi-thin slices on a 75°C oven for 15 minutes, and then add periodic acid solution in a 40°C oven for 20 minutes (periodic acid can make the polysaccharide ethylene glycol in the cells base is oxidized to dialdehyde, the periodate solution needs to be preheated, the preheating temperature is 40°C, and the time is 15min); the temperature limited in this step can speed up the combination of the subsequent dye solution and the tissue, so that the dyeing can be done in a shorter time within 2 weeks to achieve the desired stainin...

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Abstract

The invention discloses a dyeing method for tissue glycogen. The method comprises the following steps: 1, preparing sections: processing target tissues immobilized with an immobilizing solution containing glutaraldehyde and osmic acid to form the semi-thin slices; 2, carrying out periodic acid oxidation: dropwise adding a periodic acid solution onto the semi-thin slices, and oxidizing glycogen glycol groups in target tissue cells into aldehyde groups; 3, dyeing with colorless pinkish red: dyeing the periodic acid oxidized semi-thin sections by adopting a colorless pinkish red solution, and allowing the colorless pinkish red to be combined with the aldehyde groups to form a mauve color; and 4, reversely lining with toluidine blue: reversely lining the pinkish red dyed semi-thin sections with a toluidine blue solution. Compared with the prior art, the method adopting the combination of the colorless pinkish red and the toluidine blue to dye the semi-thin sections allows the presence or not of glycogen accumulation and the accurate accumulation part to be easily judged even in the presence of very less glycogen accumulation, and is very simple and reliable.

Description

technical field [0001] The invention relates to the field of biotechnology, and relates to a method for staining tissue glycogen. Background technique [0002] Glycogen storage disease caused by abnormal glucose metabolism is often seen in skeletal muscle or liver diseases. PAS staining can stain the mucopolysaccharides in the tissue into cherry red, which can show whether the glycogen has increased. [0003] Only the vacuoles in the cytoplasm can be displayed when using HE conventional tissue staining of paraffin sections, but the periodic acid-schiff (periodic acid Schiff, PAS) staining method of paraffin sections can show the accumulation of glycogen in the cytoplasm of skeletal muscle fibers. Characteristic form. PAS staining has been improved many times in paraffin sections, but there are still problems. For example, the glycogen cannot be preserved in a large amount during fixation and production, and the color after production cannot be easily and directly interprete...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N1/30
CPCG01N1/30
Inventor 侯晓涛王伶唐良玉李改岳书玲江启锋王林王家健罗丕福李晶李学锋
Owner GUANGZHOU KINGMED DIAGNOSTICS CENT
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