Zenisek David, Perrais David
CSH Protoc. 2007 Oct 1;2007:pdb.top24. doi: 10.1101/pdb.top24.
INTRODUCTIONTotal internal reflection fluorescence microscopy (TIRFM) is a powerful technique for studying events that occur near a cell surface. The technique allows selective imaging of fluorescent molecules that are closest to a high refractive index substance such as glass. TIRFM has been used to study (1) exocytosis of single synaptic vesicles stained with FM1-43 in living goldfish retinal bipolar neurons and (2) exocytosis of single dense core granules stained with neuropeptide Y-enhanced green fluorescent protein (NPY-EGFP) in living bovine chromaffin cells. This article describes the basic theory behind TIRFM and provides an overview for setting up a TIRFM system.
引言
全内反射荧光显微镜（TIRFM）是研究细胞表面附近发生的事件的一种强大技术。该技术能够对最靠近高折射率物质（如玻璃）的荧光分子进行选择性成像。TIRFM已被用于研究：（1）用FM1-43染色的单个突触小泡在活金鱼视网膜双极神经元中的胞吐作用；（2）用神经肽Y增强绿色荧光蛋白（NPY-EGFP）染色的单个致密核心颗粒在活牛嗜铬细胞中的胞吐作用。本文介绍了TIRFM背后的基本理论，并概述了TIRFM系统的搭建方法。
