Central Laser Facility, Research Complex at Harwell, Rutherford Appleton Laboratory, Harwell Oxford, Didcot, UK.
J Microsc. 2013 Oct;252(1):16-22. doi: 10.1111/jmi.12070. Epub 2013 Jul 24.
The phenomenon of total internal reflection fluorescence (TIRF) was placed in the context of optical microscopy by Daniel Axelrod over three decades ago. TIRF microscopy exploits the properties of an evanescent electromagnetic field to optically section sample regions in the close vicinity of the substrate where the field is induced. The first applications in cell biology targeted investigation of phenomena at the basolateral plasma membrane. The most notable application of TIRF is single-molecule experiments, which can provide information on fluctuation distributions and rare events, yielding novel insights on the mechanisms governing the molecular interactions that underpin many fundamental processes within the cell. This short review intends to provide a 'one stop shop' explanation of the electromagnetic theory behind the remarkable properties of the evanescent field, guide the reader through the principles behind building or choosing your own TIRF system and consider how the most popular applications of the method exploit the evanescent field properties.
三十多年前，丹尼尔·阿克塞尔罗德（Daniel Axelrod）将全内反射荧光（TIRF）现象置于光学显微镜的背景下。TIRF 显微镜利用消逝电磁场所具有的特性，对诱导该场的接近基底的样品区域进行光学切片。细胞生物学中的首批应用针对基底外侧质膜上现象的研究。TIRF 的最显著应用是单分子实验，它可以提供有关波动分布和稀有事件的信息，从而为控制分子相互作用的机制提供新的见解，这些分子相互作用是细胞内许多基本过程的基础。这篇简短的综述旨在对消逝场显著特性背后的电磁理论提供一个“一站式”的解释，引导读者了解构建或选择自己的 TIRF 系统背后的原理，并考虑该方法的最流行应用如何利用消逝场特性。
