TY - JOUR
T1 - Photocontrollable fluorescent proteins for superresolution imaging
AU - Shcherbakova, Daria M.
AU - Sengupta, Prabuddha
AU - Lippincott-Schwartz, Jennifer
AU - Verkhusha, Vladislav V.
PY - 2014/5
Y1 - 2014/5
N2 - 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.
AB - 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.
KW - EosFP
KW - PAGFP
KW - PALM
KW - PAmCherry
KW - RESOLFT
UR - http://www.scopus.com/inward/record.url?scp=84902008399&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=84902008399&partnerID=8YFLogxK
U2 - 10.1146/annurev-biophys-051013-022836
DO - 10.1146/annurev-biophys-051013-022836
M3 - Article
C2 - 24895855
AN - SCOPUS:84902008399
SN - 1936-122X
VL - 43
SP - 303
EP - 329
JO - Annual Review of Biophysics
JF - Annual Review of Biophysics
IS - 1
ER -