Comparison and optimization of special staining methods for observation of myocardial fibrosis
10.3969/j.issn.1005-4847.2024.03.008
- VernacularTitle:心肌纤维化特殊染色方法比较与优化
- Author:
Yaheng WANG
1
,
2
;
Jiaxin MA
;
Yu LEI
;
Lianfeng ZHANG
;
Dan LYU
Author Information
1. 中国医学科学院医学实验动物研究所,国家卫生健康委比较医学重点实验室,北京 100021
2. 中国医学科学院医学实验动物研究所,北京市人类重大疾病实验动物模型工程技术研究中心,北京 100021
- Keywords:
myocardial fibrosis;
special staining;
sirius red/fast green staining;
quantitative analysis
- From:
Acta Laboratorium Animalis Scientia Sinica
2024;32(3):347-354
- CountryChina
- Language:Chinese
-
Abstract:
Objective The existing dyeing methods of myocardial fibrosis were optimized to make up for the problems of missing and misreading of collagen fibers in the quantitative analysis of the current common dyeing methods of myocardial fibers,and to provide a reference for the semi-quantitative and diagnosis of myocardial fibrosis.Methods Paraffin sections of cardiac tissue were prepared using a transgenic mouse model of cardiomyopathy with a specific laboratory-constructed cTnTR141W gene mutation.Four staining method were performed for comparative observations:Masson's trichrome(Masson)staining,picrosirius red(PSR)staining,van Gieson(VG)staining,and Sirius red/fast green(SR/FG)staining.Image J 2.1.0 software was used to quantitatively compare the areas of collagen fibers.SR/FG was optimized from three aspects:dye concentration,staining time,and acid solution prestaining,and the quantitative analysis of collagen fibers was then verified.Results The collagen fiber distribution was observed by the four staining method,among which SR/FG was notable.It involved prestaining with a 0.1%Sirius red-picric acid acidic solution for 5 min,adjusting the concentration of the dye solution to 0.1%Sirius red-picric acid and 0.04%fast green mixture,and incubating the sections in the mixed staining solution for 1 h.This method exhibited the lowest incidence of missed readings and loss in determining the proportion of collagen fibers.Conclusions Compared with other traditional collagen fiber staining method,the optimized SR/FG technique described in this paper produces bright coloring of collagen fibers and myocardial tissue,obvious color contrast,and high stability,convenience,and speed.It is suitable for subsequent quantitative analysis and determination of the collagen fiber proportion.
