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A general strategy to develop cell permeable and fluorogenic probes for multi-colour nanoscopy

Authors :
Mai Tran
Julia Roberti
Elisa D’Este
Birgit Koch
Lu Wang
Kai Johnsson
Lin Xue
Publication Year :
2019
Publisher :
Cold Spring Harbor Laboratory, 2019.

Abstract

Live-cell fluorescence nanoscopy is a powerful tool to study cellular biology on a molecular scale, yet its use is held back by the paucity of suitable fluorescent probes. Fluorescent probes based on regular fluorophores usually suffer from low cell permeability and unspecific background signal. We report a general strategy to transform regular fluorophores into fluorogenic probes with excellent cell permeability and low unspecific background signal. The strategy is based on the conversion of a carboxyl group found in rhodamines and related fluorophores into an electron-deficient amide. This conversion does not affect the spectroscopic properties of the fluorophore but permits it to exist in a dynamic equilibrium between two different forms: a fluorescent zwitterion and a non-fluorescent, cell permeable spirolactam. Probes based on such fluorophores generally are fluorogenic as the equilibrium shifts towards the fluorescent form when the probe binds to its cellular targets. The resulting increase in fluorescence can be up to 1000-fold. Using this simple design principle we created fluorogenic probes in various colours for different cellular targets for wash-free, multicolour, live-cell nanoscopy. The work establishes a general strategy to develop fluorogenic probes for live-cell bioimaging.

Details

Language :
English
Database :
OpenAIRE
Accession number :
edsair.doi.dedup.....61bd83c9f5575c70f30d4f65f572696a
Full Text :
https://doi.org/10.1101/690867