Bacterial TEM: new insights from cryo-microscopy.
Some bacteria are amongst the most important model organisms for biology and medicine. Here we review how electron microscopes have been used to image bacterial cells, summarizing the technical details of the various methods, the advantages and disadvantages of each, and the major biological insights that have been obtained. Three specific example structures, "mesosomes," "cytoskeletal filaments," and "nucleoid," are used to illustrate how methodological advances have shaped our understanding of bacterial ultrastructure. Methods that involve dehydration and metal stains are widely practiced and have revealed many ultrastructural features, but they can generate misleading artifacts and have failed to preserve important structures such as the bacterial cytoskeleton. The invention of cryo-electron microscopy, which allows bacterial cells to be imaged in a frozen-hydrated, near-native state without the need for dehydration and stains, has now led to important new insights. Efforts to identify structures and localize specific proteins in cryo-EM images are summarized.
Pilhofer
Martin
M
Division of Biology, California Institute of Technology, Pasadena, California 91125, USA.
Ladinsky
Mark S
MS
McDowall
Alasdair W
AW
Jensen
Grant J
GJ
D001419:Bacteria / Q000648:ultrastructure*; D001426:Bacterial Proteins / Q000032:analysis; D020285:Cryoelectron Microscopy / Q000295:instrumentation / Q000379:methods*; D005614:Freeze Fracturing / Q000379:methods; D017110:Freeze Substitution / Q000379:methods; D007150:Immunohistochemistry / Q000379:methods; D046529:Microscopy, Electron, Transmission / Q000295:instrumentation / Q000379:methods*; D016624:Negative Staining / Q000379:methods
D001426:Bacterial Proteins