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The effect of photoswitching kinetics and labeling densities on super-resolution fluorescence imaging

Paper ID Volume ID Publish Year Pages File Format Full-Text
24077 43494 2010 7 PDF Available
Title
The effect of photoswitching kinetics and labeling densities on super-resolution fluorescence imaging
Abstract

Super-resolution fluorescence imaging methods based on reversible photoswitching of fluorophores with subsequent localization currently develop to promising tools for cellular imaging. Since most of these methods rely on the transfer of the majority of fluorophores to a non-fluorescent dark state and precise localization of separated fluorescent fluorophores, the photophysical properties of photoswitchable fluorophores have to be carefully controlled. The achievable resolution and herewith the ability to resolve a structural feature depends not only on the brightness of the fluorophores, but also on the labeling density and on the stability or lifetime of the non-fluorescent dark state. Here, we discuss how the ratio of off- and on-switching of a fluorophore affects resolution. We compare experimental data with theoretical simulations and present a strategy to customize photoswitching characteristics to achieve optimal optical resolution.

Keywords
Molecular photoswitches; Photophysics; Super-resolution imaging; dSTORM; Diffraction limit; Single-molecule based localization microscopy
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The effect of photoswitching kinetics and labeling densities on super-resolution fluorescence imaging
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Publisher
Database: Elsevier - ScienceDirect
Journal: Journal of Biotechnology - Volume 149, Issue 4, 15 September 2010, Pages 260–266
Authors
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Subjects
Physical Sciences and Engineering Chemical Engineering Bioengineering
Get Full-Text Now
Don't Miss Today's Special Offer
Price was $35.95
You save - $31
Price after discount Only $4.95
100% Money Back Guarantee
Full-text PDF Download
Online Support
Any Questions? feel free to contact us