title: Visualizing cell architecture and molecular location using soft x-ray tomography and correlated cryo-light microscopy.
abstract: Living cells are structured to create a range of microenvironments that support specific chemical reactions and processes. Understanding how cells function therefore requires detailed knowledge of both the subcellular architecture and the location of specific molecules within this framework. Here we review the development of two correlated cellular imaging techniques that fulfill this need. Cells are first imaged using cryogenic fluorescence microscopy to determine the location of molecules of interest that have been labeled with fluorescent tags. The same specimen is then imaged using soft X-ray tomography to generate a high-contrast, 3D reconstruction of the cells. Data from the two modalities are then combined to produce a composite, information-rich view of the cell. This correlated imaging approach can be applied across the spectrum of problems encountered in cell biology, from basic research to biotechnological and biomedical applications such as the optimization of biofuels and the development of new pharmaceuticals.
authors.affiliation: Department of Anatomy, University of California, San Francisco, 94158, USA.
authors.forename: Gerry
authors.initials: G
authors.lastname: McDermott
authors.forename: Mark A
authors.initials: MA
authors.lastname: Le Gros
authors.forename: Carolyn A
authors.initials: CA
authors.lastname: Larabell
mesh_terms: D000818:Animals; D004867:Equipment Design; D006801:Humans; D021621:Imaging, Three-Dimensional / Q000379:methods; D007150:Immunohistochemistry / Q000379:methods; D008853:Microscopy / Q000295:instrumentation / Q000379:methods*; D008856:Microscopy, Fluorescence / Q000295:instrumentation / Q000379:methods; D013194:Staining and Labeling / Q000379:methods; D014056:Tomography, X-Ray / Q000295:instrumentation / Q000379:methods*