活细胞中单分子的实时显视是单分子生物学的关键技术,本文针对单分子显视的光学方法做了评述.分别描述了共焦荧光显微术、荧光全内反射显微术以及荧光共振能量转移探测的技术细节, 分析了这些技术对于单分子探测所具备的优势和不足.并对单分子方法的未来发展给出预测.指出包括原子力在内的各种探测手段的联合使用和创新荧光染料技术是进一步提高分辨率的突破口.而随着高灵敏和低噪音探测器的发展,各种新方法的出现也有可能突破目前荧光染料尺度给予的分辨极限.

Due to biologic single molecular visualization in real time is a key technology to develop molecule biologics, optical detection aperture as a sensitive detection tool is reviewed in this paper. Confocal fluorescent microscopy, fluorescent total internal reflection microscopy and fluorescent resonance energy transfer detection technique are described respectively and analyzed for their advantage and shortcoming in the biologic single molecule detection. The AFM is a fine instrument capable of imaging single macromolecules with submolecular resolution. So AFM combines other optical detection methods will be a powerful tools. Furthermore, innovating technique on fluorochrome is also a rising optical resolution aperture. The resolution limitation given by the fluorochrome may be broken through with new technology, for example, reducing the fluorochrome and developing more sensitive sizeand lower noise detector, etc.