Superresolution microscopy with novel BODIPY-based fluorophores

Amy M. Bittel, Isaac S. Saldivar, Nick J. Dolman, Xiaolin Nan, Summer Gibbs

Research output: Contribution to journalArticle

3 Citations (Scopus)

Abstract

Multicolor single-molecule localization microscopy (SMLM) expands our understanding of subcellular details and enables the study of biomolecular interactions through precise visualization of multiple molecules in a single sample with resolution of ~10-20 nm. Probe selection is vital to multicolor SMLM, as the fluorophores must not only exhibit minimal spectral crosstalk, but also be compatible with the same photochemical conditions that promote fluorophore photoswitching. While there are numerous commercially available photoswitchable fluorophores that are optimally excited in the standard Cy3 channel, they are restricted to short Stokes shifts (<30 nm), limiting the number of colors that can be resolved in a single sample. Furthermore, while imaging buffers have been thoroughly examined for commonly used fluorophore scaffolds including cyanine, rhodamine, and oxazine, optimal conditions have not been found for the BODIPY scaffold, precluding its routine use for multicolor SMLM. Herein, we screened common imaging buffer conditions including seven redox reagents with five additives, resulting in 35 overall imaging buffer conditions to identify compatible combinations for BODIPY-based fluorophores. We then demonstrated that novel, photoswitchable BODIPY-based fluorophores with varied length Stokes shifts provide additional color options for SMLM using a combination of BODIPY-based and commercially available photoswitchable fluorophores.

Original languageEnglish (US)
Pages (from-to)e0206104
JournalPLoS One
Volume13
Issue number10
DOIs
StatePublished - Jan 1 2018

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Fluorophores
fluorescent dyes
Microscopy
microscopy
Microscopic examination
Buffers
Molecules
Color
Oxazines
buffers
image analysis
Rhodamines
Imaging techniques
Scaffolds
Oxidation-Reduction
color
Crosstalk
4,4-difluoro-4-bora-3a,4a-diaza-s-indacene
Single Molecule Imaging
Visualization

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)
  • Agricultural and Biological Sciences(all)

Cite this

Superresolution microscopy with novel BODIPY-based fluorophores. / Bittel, Amy M.; Saldivar, Isaac S.; Dolman, Nick J.; Nan, Xiaolin; Gibbs, Summer.

In: PLoS One, Vol. 13, No. 10, 01.01.2018, p. e0206104.

Research output: Contribution to journalArticle

Bittel, Amy M. ; Saldivar, Isaac S. ; Dolman, Nick J. ; Nan, Xiaolin ; Gibbs, Summer. / Superresolution microscopy with novel BODIPY-based fluorophores. In: PLoS One. 2018 ; Vol. 13, No. 10. pp. e0206104.
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