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Helium ion microscopy of the rodent kidney and male reproductive tract
Author(s) -
Paunescu Teodor G.,
Huynh Chuong,
Lechner Lorenz,
Goetze Bernhard,
Stern Lewis,
Breton Sylvie,
Brown Dennis
Publication year - 2013
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.27.1_supplement.683.1
Subject(s) - epididymis , stereocilia (inner ear) , microbiology and biotechnology , microscopy , cilium , nuclear membrane , ultrastructure , biophysics , chemistry , kidney , biology , sperm , anatomy , pathology , cytoplasm , medicine , hair cell , botany , endocrinology , cochlea
We investigated the epithelia of the rat and mouse kidney and epididymis by helium ion microscopy (HIM), a novel technology that uses a scanning beam of He+ ions to produce nanometer resolution images of uncoated biological samples. Various tissue fixation, sectioning, and dehydration methods were evaluated for their ability to preserve tissue architecture. Samples were subjected to critical point drying and HIM, allowing the visualization of fine cellular and plasma membrane details, including depressions suggesting endo‐ or exocytotic events, cilia, and microvesicles. In the kidney, membranous nanoprojections on podocytes and pores within the filtration slit diaphragm were seen in the glomerulus, and extensive apical microplicae of the intercalated cells in the collecting duct. In the epididymis, stereocilia in principal cells and microplicae in clear cells are clearly distinguishable, and high resolution imaging of sperm reveals their close interaction with the epithelium. Colloidal gold probes were used in conjunction with HIM, allowing us to localize membrane proteins. In conclusion, this technology represents a major breakthrough in imaging of the topographical ultrastructure of biological samples and should provide significant advances in our understanding of cell surface structures and membrane organization. Funding sources: NIH DK42956, DK97124, and HD40793; Carl Zeiss Microscopy, LLC.