Mechanistic investigation of mEos4b reveals a strategy to reduce track interruptions in sptPALM.

Authors:
Dr. Virgile Adam, PhD
Dr. Virgile Adam, PhD
CNRS / Institute for Structural Biology (IBS)
Research scientist
single molecule fluorescence microscopy, fluorescent protein engineering, X-ray crystallography
Grenoble | France

Nat Methods 2019 08 8;16(8):707-710. Epub 2019 Jul 8.

University Grenoble Alpes, CEA, CNRS, IBS, Grenoble, France.

Green-to-red photoconvertible fluorescent proteins repeatedly enter dark states, causing interrupted tracks in single-particle-tracking localization microscopy (sptPALM). We identified a long-lived dark state in photoconverted mEos4b that results from isomerization of the chromophore and efficiently absorbs cyan light. Addition of weak 488-nm light swiftly reverts this dark state to the fluorescent state. This strategy largely eliminates slow blinking and enables the recording of longer tracks in sptPALM with minimum effort.

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http://dx.doi.org/10.1038/s41592-019-0462-3DOI Listing
August 2019
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