Veerapathiran Sapthaswaran, Wohland Thorsten
Department of Biological Sciences and NUS Centre for Bio-Imaging Sciences, National University of Singapore, 14 Science Drive 4, Singapore 117557, Singapore.
J Biosci. 2018 Jul;43(3):541-553.
Advanced fluorescence techniques, commonly known as the F-techniques, measure the kinetics and the interactions of biomolecules with high sensitivity and spatiotemporal resolution. Applications of the F-techniques, which were initially limited to cells, were further extended to study in vivo protein organization and dynamics in whole organisms. The integration of F-techniques with multi-photon microscopy and light-sheet microscopy widened their applications in the field of developmental biology. It became possible to penetrate the thick tissues of living organisms and obtain good signal-to-noise ratio with reduced photo-induced toxicity. In this review, we discuss the principle and the applications of the three most commonly used F-techniques in developmental biology: Fluorescence Recovery After Photo-bleaching (FRAP), Fo¨ rster Resonance Energy Transfer (FRET), and Fluorescence Correlation and Cross-Correlation Spectroscopy (FCS and FCCS).
先进的荧光技术，通常称为F技术，能够以高灵敏度和时空分辨率测量生物分子的动力学和相互作用。F技术最初仅应用于细胞，后来其应用范围进一步扩展到研究整个生物体中的体内蛋白质组织和动力学。F技术与多光子显微镜和光片显微镜的结合拓宽了它们在发育生物学领域的应用。穿透活生物体的厚组织并获得具有降低的光诱导毒性的良好信噪比成为可能。在本综述中，我们讨论发育生物学中最常用的三种F技术的原理和应用：光漂白后荧光恢复（FRAP）、Förster共振能量转移（FRET）以及荧光相关和交叉相关光谱（FCS和FCCS）。
