Imaging Exocytosis with Total Internal Reflection Microscopy (TIRFM).
Cold Spring Harb Protoc. 2007-10-01; 2007(10): pdb.prot4863
DOI: 10.1101/pdb.prot4863
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1. CSH Protoc. 2007 Oct 1;2007:pdb.prot4863. doi: 10.1101/pdb.prot4863.
Imaging Exocytosis with Total Internal Reflection Microscopy (TIRFM).
Zenisek D, Perrais D.
INTRODUCTIONAlthough electrophysiological techniques such as membrane capacitance
measurements, electrochemical detection, and post-synaptic recordings are
powerful ways of studying exocytosis, information concerning any steps prior to
vesicle fusion must be inferred indirectly. Total internal reflection
fluorescence microscopy (TIRFM) is a powerful technique for studying events that,
like exocytosis, occur near a cell surface. The technique allows selective
imaging of fluorescent molecules that are closest to a high refractive index
substance such as glass. In this protocol, TIRFM is used to investigate the steps
leading up to vesicle fusion in both retinal bipolar neurons and chromaffin cells
by directly imaging synaptic vesicles and dense core granules prior to and
including exocytosis.
PMID: 21356953