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Characterization of intact subcellular bodies in whole bacteria by cryo‐electron tomography and spectroscopic imaging
Author(s) -
COMOLLI L. R.,
KUNDMANN M.,
DOWNING K. H.
Publication year - 2006
Publication title -
journal of microscopy
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.569
H-Index - 111
eISSN - 1365-2818
pISSN - 0022-2720
DOI - 10.1111/j.1365-2818.2006.01597.x
Subject(s) - caulobacter crescentus , bacteria , cryo electron tomography , chemistry , electron microscope , biophysics , electron tomography , characterization (materials science) , tomography , biochemistry , biology , materials science , cell , nanotechnology , physics , optics , scanning transmission electron microscopy , genetics , cell cycle
Summary We illustrate the combined use of cryo‐electron tomography and spectroscopic difference imaging in the study of subcellular structure and subcellular bodies in whole bacteria. We limited our goal and focus to bodies with a distinct elemental composition that was in a sufficiently high concentration to provide the necessary signal‐to‐noise level at the relatively large sample thicknesses of the intact cell. This combination proved very powerful, as demonstrated by the identification of a phosphorus‐rich body in Caulobacter crescentus . We also confirmed the presence of a body rich in carbon, demonstrated that these two types of bodies are readily recognized and distinguished from each other, and provided, for the first time to our knowledge, structural information about them in their intact state. In addition, we also showed the presence of a similar type of phosphorus‐rich body in Deinococcus grandis , a member of a completely unrelated bacteria genus. Cryo‐electron microscopy and tomography allowed the study of the biogenesis and morphology of these bodies at resolutions better than 10 nm, whereas spectroscopic difference imaging provided a direct identification of their chemical composition.