Far-red photoactivatable BODIPYs for the super-resolution imaging of live cells

Yang Zhang; Sicheng Tang; Laura Ravelo; Janet Cusido; Françisco M Raymo

doi:10.1016/bs.mie.2020.04.026

Far-red photoactivatable BODIPYs for the super-resolution imaging of live cells

Methods Enzymol. 2020:640:131-147. doi: 10.1016/bs.mie.2020.04.026. Epub 2020 May 18.

Authors

Yang Zhang¹, Sicheng Tang², Laura Ravelo², Janet Cusido³, Françisco M Raymo⁴

Affiliations

¹ Laboratory for Molecular Photonics, Department of Chemistry, University of Miami, Coral Gables, FL, United States; Department of Biomedical Engineering, Northwestern University, Evanston, IL, United States.
² Laboratory for Molecular Photonics, Department of Chemistry, University of Miami, Coral Gables, FL, United States.
³ Laboratory for Molecular Photonics, Department of Chemistry, University of Miami, Coral Gables, FL, United States; Department of Math and Natural Sciences, Miami Dade College-Eduardo J. Padron Campus, Miami, FL, United States.
⁴ Laboratory for Molecular Photonics, Department of Chemistry, University of Miami, Coral Gables, FL, United States. Electronic address: fraymo@miami.edu.

PMID: 32560795
DOI: 10.1016/bs.mie.2020.04.026

Abstract

The identification of viable designs to construct switchable fluorescent probes and operate them in the interior of live cells is essential to allow the acquisition of SMLM images and permit the visualization of cellular components with sub-diffraction resolution. Our laboratories developed a mechanism to switch the fluorescence of BODIPY chromophores with the photoinduced cleavage of oxazine heterocycles under mild 405-nm illumination. With appropriate structural modifications, these switchable molecules can be engineered to immobilize covalently on large biomolecules within lysosomal compartments of live COS-7 cells and produce bright far-red fluorescence with optimal contrast upon activation. Such a combination of properties permits the acquisition of PALM images of the labeled organelles with localization precision of ca. 15nm. This article reports the experimental protocols for the synthesis of and live-cell labeling with these compounds as well as for the reconstruction of super-resolution images of the resulting biological preparations.

Keywords: BODIPY; Oxazines; PALM; Photoactivatable fluorophores; Super-resolution imaging.

Publication types

Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

Animals
Boron Compounds*
COS Cells
Chlorocebus aethiops
Fluorescent Dyes*
Microscopy, Fluorescence

Substances

4,4-difluoro-4-bora-3a,4a-diaza-s-indacene
Boron Compounds
Fluorescent Dyes