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Cross-talk-free multi-color STORM imaging using a single fluorophore.

ABSTRACT: Multi-color stochastic optical reconstruction microscopy (STORM) is routinely performed; however, the various approaches for achieving multiple colors have important caveats. Color cross-talk, limited availability of spectrally distinct fluorophores with optimal brightness and duty cycle, incompatibility of imaging buffers for different fluorophores, and chromatic aberrations impact the spatial resolution and ultimately the number of colors that can be achieved. We overcome these complexities and develop a simple approach for multi-color STORM imaging using a single fluorophore and sequential labelling. In addition, we present a simple and versatile method to locate the same region of interest on different days and even on different microscopes. In combination, these approaches enable cross-talk-free multi-color imaging of sub-cellular structures.

SUBMITTER: Tam J

PROVIDER: S-EPMC4084994 | biostudies-literature | 2014

REPOSITORIES: biostudies-literature

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Cross-talk-free multi-color STORM imaging using a single fluorophore.

Tam Johnny J Cordier Guillaume Alan GA Borbely Joseph Steven JS Sandoval Álvarez Angel A Lakadamyali Melike M

PloS one 20140707 7

Multi-color stochastic optical reconstruction microscopy (STORM) is routinely performed; however, the various approaches for achieving multiple colors have important caveats. Color cross-talk, limited availability of spectrally distinct fluorophores with optimal brightness and duty cycle, incompatibility of imaging buffers for different fluorophores, and chromatic aberrations impact the spatial resolution and ultimately the number of colors that can be achieved. We overcome these complexities an ...[more]

PMID: 25000286

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