隐失波荧光显微镜及其在植物细胞生物学中的应用

Evanescent wave microscopy and its application in plant cell biology

隐失波荧光显微术(evanescent wave fluorescence microscopy,EWFM)是一种对细胞膜附近的生物学过程进行选择性观测的光学显微成像技术,该技术是基于全内反射原理,即当光在生物样品表面产生全内反射时,虽然在样品表面未被光直接照亮,但它仍会产生一种隐失波,这种隐失波却有足够能量将样品表面100～200 nm范围内的荧光基团激发。由于这一特性使得隐失波荧光显微镜能对靠近细胞膜的生物学过程,如胞吞胞吐、单个分子与细胞膜的结合和解离,以及膜上受体的动态变化过程等进行更好的观测。本文简略地介绍了隐失波荧光显微术的原理,同时对其在细胞生物学,特别是在植物细胞生物学中的应用作了初步总结。随着隐失波荧光显微镜的商业化及其观测技术的不断改进,它将在生命科学领域的研究中发挥越来越大的作用。

Evanescent wave microscopy is an elegant optical technique which can conduct the selective imaging of molecular events very close to the plasma membrane.This state-of-the-art optical approach to biological process based on the evanescent field that is generated beyond a reflecting surface at the interface between two transparent materials of different refractive indexes.Although there is no direct illumination of the sample with the laser light,the evanescent field can penetrate 100 ～ 200 nm into the sample near the coverslip.These attributes make evanescent wave microscopy ideal for selectively imaging membrane-associated processes such as endocytosis,exocytosis,trafficking of molecules to and from the cellular plasma membrane,the dynamics of the membrane-bound receptors.This review will mainly focus on the principle of evanescent wave microscopy and its application to cell biology,especially to the plant cell biology.As commercial evanescent wave microscopes starts to become an standard equipment in laboratories and innovative improvements extend the boundaries of this technique,it is clear that evanescent wave microscopy faces a bright future.