Application of en bloc Staining Technique in Three-Dimensional Ultrastructure Reconstruction of Biological Samples

YAO Xiaomin^1,2, LI Mingyang¹, LI Changyuan¹, YUAN Xiaoyan^1,2, SHEN Weiwei^1,2, LI Xiaojuan^1,2*

(¹Key Laboratory of Genetics and Breeding in Forest Trees and Ornamental Plants, Ministry of Education, College of Bioscience and Bioengineering, Beijing Forestry University, Beijing 100083, China; ²Institute of Tree Development and Genome Editing, Beijing Forestry University, Beijing 100083, China)

Abstract:

Investigating the detailed information of localization and interaction of bio-macromolecules and organelles will provide valuable insights into the understanding of vital life processes. Therefore, three-dimensional reconstructions of the ultrastructure of large-volume biological samples by volume electron microscopy have promoted the research of cell biology and neurobiology. Since the biological sample itself only behaves weak electron contrast, the detailed ultrastructure of the sample under electron microscope usually presented unclear. The staining techniques can effectively increase the electron scattering difference of the sample, therefore greatly improves the image quality. En bloc staining has been successfully used in studying the ultrastructure of large-volume samples. This review summarizes the specimen preparation process, staining method and the characteristics of different bridging agents and en bloc staining agents, in order to provide some insights for the application and development of en bloc staining technology.

CSTR: 32200.14.cjcb.2022.11.0010