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Cold Spring Harb Protoc
2011[Dec]; 2011
(12
): 1498-506
PMID22135665
show ga
Gold nanoparticles, like single fluorophores, can be used to locate single
molecules with nanometer accuracy. Unlike an optical trap, the gold particle
label does not exert an external load, which is important for studying diffusive
processes. Thus, a gold particle can be used analogously to a single fluorophore,
providing similar information but with submillisecond time resolution. The
features of gold-nanoparticle tracking (high temporal resolution, small label
size, and lack of applied force) facilitate the characterization of structural
properties of short-lived intermediates, as shown by our work with myosin V. This
protocol provides details for gold-nanoparticle-tracking experiments, including
flow cell construction, microscopy, and data analysis, along with a brief outline
of actin and myosin preparation. Although details particular to our experiment
are given, the approach should be generally applicable.
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