中国实验动物学报2024，Vol.32Issue(3)：347-354,8.DOI:10.3969/j.issn.1005-4847.2024.03.008
心肌纤维化特殊染色方法比较与优化
Comparison and optimization of special staining methods for observation of myocardial fibrosis
摘要
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.关键词
心肌纤维化/特殊染色方法/天狼星红-固绿/定量分析Key words
myocardial fibrosis/special staining/sirius red/fast green staining/quantitative analysis分类
生物科学引用本文复制引用
王亚恒,马嘉昕,雷雨,张连峰,吕丹..心肌纤维化特殊染色方法比较与优化[J].中国实验动物学报,2024,32(3):347-354,8.基金项目
中国医学科学院医学与健康科技创新工程(2022-I2M-1-020),中央高校基本科研业务费(3332023054). Funded by Chinese Academy of Medical Sciences(CAMS)Innovation Fund for Medical Sciences(2022-I2M-1-020),Special Research Fund for Central Universities,Peking Union Medical College(PUMC)(3332023054). （2022-I2M-1-020）
