Fluorescence Microscopy and Thin-Section Electron Microscopy
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Intracellular structures in Pichia pastoris can be visualized by the complementary methods of fluorescence microscopy and electron microscopy. An improved immunofluorescence
protocol yields better optics and more reliable antigen preservation than conventional methods. As an alternative to immunofluorescence,
if a protein of interest is fused to GFP or another fluorescent tag, the cells can be fixed and viewed directly. For higher-resolution
studies of organelle morphology, thin-section electron microscopy of permanganate-fixed cells yields good preservation of
intracellular membranes.
Affiliation(s): (2) Department of Molecular Genetics and Cell Biology, The University of Chicago, Chicago, IL
Book Title: Pichia Protocols
Series: Methods in Molecular Biology | Volume: 389 | Pub. Date: Aug-08-2007 | Page Range: 251-259 | DOI: 10.1007/978-1-59745-456-8_18
Subject: Biotechnology
Key Words: Immunofluorescence - fluorescence microscopy - electron microscopy - yeast - Pichia pastoris - fluorescent proteins - permanganate fixation
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