Journal
BIOPHYSICAL JOURNAL
Volume 96, Issue 2, Pages L22-L24Publisher
CELL PRESS
DOI: 10.1016/j.bpj.2008.11.002
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Funding
- Health Research Council of New Zealand
- Auckland University Research Committee
- Emma Kay
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We show that high quantum efficiency fluorophores can exhibit reversible photobleaching. This observation provides the basis for an imaging technique we call reversible photobleaching microscopy. We demonstrate applicability of this technique using antibody labeled biological samples in standard aqueous (or glycerol based) media to produce far-field images at similar to 30 nm resolution. Our novel method relies on intense illumination to reversibly induce a very long-lived (>10 s) dark state from which single fluorochromes slowly return stochastically. As in other localization microscopy methods, reversible photobleaching microscopy localizes single fluorochromes, but has the advantage that specialized photoactivatible and photo-switchable molecules or special immersion/embedding media are not required.
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