High-pressure freezing, cellular tomography, and structural cell biology.
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 temperature 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.
McDonald
Kent L
KL
Electron Microscope Laboratory, University of California, Berkeley, USA.
Auer
Manfred
M
D000818:Animals; D002477:Cells / Q000648:ultrastructure*; D015925:Cryopreservation / Q000379:methods*; D017110:Freeze Substitution / Q000379:methods; D005615:Freezing; D021621:Imaging, Three-Dimensional; D008854:Microscopy, Electron / Q000639:trends*; D011312:Pressure; D014054:Tomography