Advanced fluorescence techniques, commonly known as the F-techniques, measure the kinetics and the interactions ofbiomolecules with high sensitivity and spatiotemporal resolution. Applications of the F-techniques, which were initiallylimited to cells, were further extended to study in vivo protein organization and dynamics in whole organisms. The integrationof F-techniques with multi-photon microscopy and light-sheet microscopy widened their applications in the field of developmentalbiology. It became possible to penetrate the thick tissues of living organisms and obtain good signal-to-noise ratiowith reduced photo-induced toxicity. In this review, we discuss the principle and the applications of the three most commonlyused F-techniques in developmental biology: Fluorescence Recovery After Photo-bleaching (FRAP), Fo¨ rster ResonanceEnergy Transfer (FRET), and Fluorescence Correlation and Cross-Correlation Spectroscopy (FCS and FCCS).
Volume 45, 2020
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