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Micro‐CT scouting for transmission electron microscopy of human tissue specimens
Authors:A.G. MORALES  E.S. STEMPINSKI  X. XIAO  A. PATEL  A. PANNA  K.N. OLIVIER  P.J. MCSHANE  C. ROBINSON  A.J. GEORGE  D.R. DONAHUE  P. CHEN  H. WEN
Affiliation:1. National Heart Lung and Blood Institute, National Institutes of Health, Bethesda, Maryland, U.S.A.;2. Advanced Photon Source, Argonne National Laboratory, Lemont, Illinois, U.S.A.;3. National Institute of Neurological Disorders and Stroke, National Institutes of Health, Bethesda, Maryland, U.S.A.
Abstract:Transmission electron microscopy (TEM) provides sub‐nanometre‐scale details in volumetric samples. Samples such as pathology tissue specimens are often stained with a metal element to enhance contrast, which makes them opaque to optical microscopes. As a result, it can be a lengthy procedure to find the region of interest inside a sample through sectioning. We describe micro‐CT scouting for TEM that allows noninvasive identification of regions of interest within a block sample to guide the sectioning step. In a tissue pathology study, a bench‐top micro‐CT scanner with 10 μm resolution was used to determine the location of patches of the mucous membrane in osmium‐stained human nasal scraping samples. Once the regions of interest were located, the sample block was sectioned to expose that location, followed by ultra‐thin sectioning and TEM to inspect the internal structure of the cilia of the membrane epithelial cells with nanometre resolution. This method substantially reduced the time and labour of the search process from typically 20 sections for light microscopy to three sections with no added sample preparation.
Keywords:bench‐top micro‐CT scanner  micro‐CT scouting  three‐dimensional visualization  transmission electron microscopy
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