Jameson David M, James Nicholas G, Albanesi Joseph P
Department of Cell and Molecular Biology, John A. Burns School of Medicine, University of Hawaii, Honolulu, Hawaii, USA.
Methods Enzymol. 2013;519:87-113. doi: 10.1016/B978-0-12-405539-1.00003-8.
Communication between cells and their environment, including other cells, is often mediated by cell surface receptors. Fluorescence methodologies are among the most important techniques used to study receptors and their interactions, and in the past decade, fluorescence fluctuation spectroscopy (FFS) approaches have been increasingly utilized. In this overview, we illustrate how diverse FFS approaches have been used to elucidate important aspects of receptor systems, including interactions of receptors with their ligands and receptor oligomerization and clustering. We also describe the most popular methods used to introduce fluorescent moieties into the biological systems. Finally, specific attention will be given to cell maintenance and transfection strategies especially as related to microscopy studies.
细胞与其环境(包括其他细胞)之间的通讯通常由细胞表面受体介导。荧光方法是用于研究受体及其相互作用的最重要技术之一,在过去十年中,荧光涨落光谱法(FFS)已得到越来越广泛的应用。在本综述中,我们阐述了如何运用多种FFS方法来阐明受体系统的重要方面,包括受体与其配体的相互作用、受体寡聚化和聚集。我们还描述了将荧光基团引入生物系统的最常用方法。最后,将特别关注细胞维持和转染策略,尤其是与显微镜研究相关的策略。