Photocontrollable fluorescent proteins for superresolution imaging

Daria M. Shcherbakova, Prabuddha Sengupta, Jennifer Lippincott-Schwartz, Vladislav V. Verkhusha

Research output: Contribution to journalArticlepeer-review

172 Scopus citations


Superresolution fluorescence microscopy permits the study of biological processes at scales small enough to visualize fine subcellular structures that are unresolvable by traditional diffraction-limited light microscopy. Many superresolution techniques, including those applicable to live cell imaging, utilize genetically encoded photocontrollable fluorescent proteins. The fluorescence of these proteins can be controlled by light of specific wavelengths. In this review, we discuss the biochemical and photophysical properties of photocontrollable fluorescent proteins that are relevant to their use in superresolution microscopy. We then describe the recently developed photoactivatable, photoswitchable, and reversibly photoswitchable fluorescent proteins, and we detail their particular usefulness in single-molecule localization-based and nonlinear ensemble-based superresolution techniques. Finally, we discuss recent applications of photocontrollable proteins in superresolution imaging, as well as how these applications help to clarify properties of intracellular structures and processes that are relevant to cell and developmental biology, neuroscience, cancer biology and biomedicine.

Original languageEnglish (US)
Pages (from-to)303-329
Number of pages27
JournalAnnual Review of Biophysics
Issue number1
StatePublished - May 2014


  • EosFP
  • PALM
  • PAmCherry

ASJC Scopus subject areas

  • Biophysics
  • Structural Biology
  • Bioengineering
  • Biochemistry
  • Cell Biology


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