Hedde Per Niklas, Nienhaus Gerd Ulrich
Institute of Applied Physics and Center for Functional Nanostructures (CFN), Karlsruhe Institute of Technology (KIT), 76128, Karlsruhe, Germany.
Department of Physics, University of Illinois at Urbana-Champaign, Urbana, IL, 61801, USA.
Biophys Rev. 2010 Dec;2(4):147-158. doi: 10.1007/s12551-010-0037-0. Epub 2010 Sep 8.
Visualization of subcellular structures and their temporal evolution is of utmost importance to understand a vast range of biological processes. Optical microscopy is the method of choice for imaging live cells and tissues; it is minimally invasive, so processes can be observed over extended periods of time without generating artifacts due to intense light irradiation. The use of fluorescence microscopy is advantageous because biomolecules or supramolecular structures of interest can be labeled specifically with fluorophores, so the images reveal information on processes involving only the labeled molecules. The key restriction of optical microscopy is its moderate resolution, which is limited to about half the wavelength of light (∼200 nm) due to fundamental physical laws governing wave optics. Consequently, molecular processes taking place at spatial scales between 1 and 100 nm cannot be studied by regular optical microscopy. In recent years, however, a variety of super-resolution fluorescence microscopy techniques have been developed that circumvent the resolution limitation. Here, we present a brief overview of these techniques and their application to cellular biophysics.
可视化亚细胞结构及其时间演变对于理解众多生物过程至关重要。光学显微镜是对活细胞和组织进行成像的首选方法；它具有微创性，因此可以长时间观察过程，而不会因强光照射产生伪影。荧光显微镜的使用具有优势，因为感兴趣的生物分子或超分子结构可以用荧光团特异性标记，所以图像揭示了仅涉及标记分子的过程的信息。光学显微镜的关键限制是其适度的分辨率，由于波动光学的基本物理定律，其分辨率限制在光波长的大约一半（约200纳米）。因此，在1到100纳米空间尺度上发生的分子过程无法通过常规光学显微镜进行研究。然而，近年来，已经开发出多种超分辨率荧光显微镜技术，这些技术规避了分辨率限制。在这里，我们简要概述这些技术及其在细胞生物物理学中的应用。
