Wang Yingxiao, Shyy John Y-J, Chien Shu
Department of Bioengineering and Beckman Institute for Advanced Science and Technology, University of Illinois, Urbana-Champaign, Urbana, IL 61801, USA.
Annu Rev Biomed Eng. 2008;10:1-38. doi: 10.1146/annurev.bioeng.010308.161731.
Fluorescence proteins (FPs) have been widely used for live-cell imaging in the past decade. This review summarizes the recent advances in FP development and imaging technologies using FPs to monitor molecular localization and activities and gene expressions in live cells. We also discuss the utilization of FPs to develop molecular biosensors and the principles and application of advanced technologies such as fluorescence resonance energy transfer (FRET), fluorescence recovery after photobleaching (FRAP), fluorescence lifetime imaging microscopy (FLIM), and chromophore-assisted light inactivation (CALI). We present examples of the application of FPs and biosensors to visualize mechanotransduction events with high spatiotemporal resolutions in live cells. These live-cell imaging technologies, which represent a frontier area in biomedical engineering, can shed new light on the mechanisms regulating mechanobiology at cellular and molecular levels in normal and pathophysiological conditions.
在过去十年中,荧光蛋白(FPs)已广泛用于活细胞成像。本综述总结了荧光蛋白开发和成像技术的最新进展,这些技术利用荧光蛋白监测活细胞中的分子定位、活性和基因表达。我们还讨论了利用荧光蛋白开发分子生物传感器以及诸如荧光共振能量转移(FRET)、光漂白后荧光恢复(FRAP)、荧光寿命成像显微镜(FLIM)和发色团辅助光灭活(CALI)等先进技术的原理和应用。我们展示了荧光蛋白和生物传感器在以高时空分辨率可视化活细胞中机械转导事件方面的应用实例。这些活细胞成像技术代表了生物医学工程的前沿领域,能够为正常和病理生理条件下细胞和分子水平上调节力学生物学的机制提供新的见解。