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

Yang Zhang, Sicheng Tang, Laura Ravelo, Janet Cusido, Françisco M. Raymo*

*Corresponding author for this work

Research output: Chapter in Book/Report/Conference proceedingChapter

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. 15 nm. 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.

Original languageEnglish (US)
Title of host publicationChemical Tools for Imaging, Manipulating, and Tracking Biological Systems
Subtitle of host publicationDiverse Methods Based on Optical Imaging and Fluorescence
EditorsDavid M. Chenoweth
PublisherAcademic Press Inc
Pages131-147
Number of pages17
ISBN (Print)9780128211533
DOIs
StatePublished - 2020

Publication series

NameMethods in Enzymology
Volume640
ISSN (Print)0076-6879
ISSN (Electronic)1557-7988

Keywords

  • BODIPY
  • Oxazines
  • PALM
  • Photoactivatable fluorophores
  • Super-resolution imaging

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology

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