井冈山大学学报（自然科学版）2026，Vol.47Issue(1)：60-67,8.DOI:10.3969/j.issn.1674-8085.2026.01.008
基于块染技术的电镜生物样本免切片染色制备方法探究
Exploration of sectioning-free staining preparation methods for biological samples in electron microscopy based on en-block staining techniques
摘要
Abstract
The appropriate block staining method is crucial for ensuring the quality of biological electron microscopy imaging.Based on conventional biological sample block staining,this study systematically investigated 13 direct block staining methods for biological samples that can be observed with a transmission electron microscope(TEM)before embedding,using mouse kidneys as a model.The results showed that the observation of ultrastructure was affected by different staining reagents and staining sequences,compared with the conventional uranium block staining method.Among them,the samples prepared by the osmium tetroxide-thiocarbohydrazide-osmium tetroxide(OTO method),the osmium tetroxide-tannic acid-tannic acid-osmium tetroxide(OTTO method),and the potassium ferrocyanide-osmium tetroxide-thiocarbohydrazide-osmium tetroxide(ROTO method)showed improved structural clarity and image contrast of organelles such as mitochondria.The staining sequence of heavy metals lead and uranium was found to affect the clarity of mitochondrial cristae structure.The optimized potassium ferrocyanide-osmium tetroxide-tannic acid-tannic acid-osmium tetroxide(ROT(T)O method),when combined with lead and uranium double block staining,not only increased the sample contrast,but also more effectively maintained the cytoskeleton and cell membrane structure,thereby achieving the best imaging effect.关键词
生物样本/块染色/透射电镜/铅、铀双重块染色Key words
biological specimens/en-block staining/transmission electron microscope/lead and uranium double en-block staining分类
生物科学引用本文复制引用
蒲琪,范京川,黄戎娟,位全芳..基于块染技术的电镜生物样本免切片染色制备方法探究[J].井冈山大学学报（自然科学版）,2026,47(1):60-67,8.基金项目
重庆市教委雏鹰计划研究项目(CY240403) （CY240403）
重庆市沙坪坝区2024年技术创新项目(2024018) （2024018）
重庆医科大学教改项目(JY20220311) （JY20220311）
