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

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...

Full description

Saved in:
Bibliographic Details
Published in:Nature methods Vol. 16; no. 8; pp. 707 - 710
Main Authors: De Zitter, Elke, Thédié, Daniel, Mönkemöller, Viola, Hugelier, Siewert, Beaudouin, Joël, Adam, Virgile, Byrdin, Martin, Van Meervelt, Luc, Dedecker, Peter, Bourgeois, Dominique
Format: Journal Article
Language:English
Published: United States Nature Publishing Group 01.08.2019
Subjects:
Blinking
Chromophores
Fluorescence
Isomerization
Lasers
Light
Localization
Microscopy
Proteins
Recording
Tracking
ISSN:1548-7091, 1548-7105, 1548-7105
Online Access:Get full text
Tags: Add Tag
No Tags, Be the first to tag this record!
Description
Summary: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.
Bibliography:ObjectType-Article-1
SourceType-Scholarly Journals-1
ObjectType-Feature-2
content type line 14
content type line 23
ISSN:1548-7091
1548-7105
1548-7105
DOI:10.1038/s41592-019-0462-3