Open AccessHigh-Pressure Freezing, Cellular Tomography, and Structural Cell Biology
Author(s) -
Kent McDonald,
Manfred Auer
Publication year - 2006
Publication title -
biotechniques/biotechniques
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.617
H-Index - 131
eISSN - 1940-9818
pISSN - 0736-6205
DOI - 10.2144/000112226
Subject(s) - ultrastructure , electron microscope , electron tomography , microscopy , resolution (logic) , biophysics , microbiology and biotechnology , cryo electron microscopy , nanotechnology , biology , materials science , physics , optics , computer science , anatomy , scanning transmission electron microscopy , artificial intelligence
Structural cell biology, which we define as electron microscopic analysis of intact cells, suffered a loss of interest and activity following the advances in light microscopy beginning in the 1990s. Interestingly, it is the wealth of detailed observation in the light microscope that is one of the driving forces for the current renewed interest in electron microscopy (EM). A great many cellular details are simply beyond the resolving power of the light microscope. In this article, we describe how electron microscopists are responding to the demands for better preservation of cells and for ways to view cell ultrastructure in three dimensions at high resolution. We discuss how low tem perature methods, especially high-pressure freezing and freeze substitution, reduce the artifacts of conventional EM specimen preparation. We also give a brief introduction to cellular electron tomography, a powerful analytical method that can give near-atomic resolution of cell ultrastructure in three-dimensional (3-D) models.