Technical advances in intracellular detection using immuno-gold particles: simple cryofixation with metal contact quick freezing

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Title
Technical advances in intracellular detection using immuno-gold particles: simple cryofixation with metal contact quick freezing
Author(s)
C Song; Ju Huck Lee; S Jun; J M Chung; J Hyun; H S Jung
Bibliographic Citation
Journal of Nanoscience and Nanotechnology, vol. 16, no. 5, pp. 4593-4597
Publication Year
2016
Abstract
The preparation of biological specimens using cryofixation techniques ensures excellent visibility of intracellular structures and preserves the antigenic sites of subcellular molecules. Hence, cryofixation is an effective method of preparing samples for analyses using antibodies conjugated to gold nanoparticles that are designed to detect the localization of specific target molecules within cells. However, cryofixation cannot be utilized easily because it requires expensive equipment and skilled technologists, resulting in a high level of expense for researchers. Here, we describe a simple technical approach to cryofixation that uses metal contact quick freezing followed by a modified freeze substitution technique and immuno-gold labeling electron microscopy. Micrograph images of cells prepared using this modified cryofixation method demonstrated its superiority over chemical fixation for high contrast visualization of the morphologies of cellular components and preservation of antigenicity for immuno-gold labeling. This report provides valuable technical information related to the advancement of metal contact quick freezing techniques, which can be used to visualize biomedical events of interest in an easy, simple, and rapid manner.
Keyword
Biomedical imagingElectron microscopyImmuno-gold labelingMetal contact quick freezingNanoparticles
ISSN
1533-4880
Publisher
Amer Scientific Publishers
DOI
http://dx.doi.org/10.1166/jnn.2016.11002
Type
Article
Appears in Collections:
Jeonbuk Branch Institute > Biological Resource Center > 1. Journal Articles
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