全内反射荧光显微术

全内反射荧光显微技术是当今世界上最具前途的新型生物光学显微技术之一，可以用来实现对单个荧光分子的直接探测。它利用全内反射产生的隐失波照明样品，使照明区域限定在样品表面的一薄层范围内，因此具有其它光学成像技术无法比拟的高的信噪比和对比度。近上来，已被生物物理学家们广泛应用于单分子的荧光成像中。本文系统的介绍了全内反射荧光显微技术的原理。国内外的发展和现状及其在生物学上的应用，并对其未来做了展望。

TOTAL INTERNAL REFLECTION FLUORESCENCE MICROSCOPY

The total internal reflection fluorescence microscopy is one of the most promising bio optical techniques in the world, which has been used to observe single fluorescent molecule directly. This specialized technique specifically illuminates sample with evanescent wave created by total internal reflection, and only a few hundred nanometers depth of sample surface should be excited. As a result, it provides the highest signal to noise ratio and contrast that is unmatched by any other light microscopy technique. Recently, it is gaining popularity with biophysicist to observe single fluorescent molecules. In this paper, the principle of the total internal reflection fluorescence microscopy and the process at home and abroad are systematically reviewed, as well as the application in cell biology is introduced. Future prospect of this technology are also discussed.