Three-dimensional imaging of single molecules solvated in pores of poly(acrylamide) gels
Three-dimensional imaging of single molecules solvated in pores of poly(acrylamide) gels
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DOI：
10.1126/science.274.5289.966
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发表时间：
1996-11-08
期刊：
影响因子：
56.9
通讯作者：
Moerner, WE
中科院分区：
文献类型：
--
作者：
Dickson, RM;Norris, DJ;Moerner, WE
通过焦油场显微镜，在聚丙烯酰胺凝胶的水填充孔中观察到单个荧光分子和单个单标记蛋白。布朗运动显着减少了凝胶框架，从而使单荧光团在水性环境中的扩展研究。一个高度轴向依赖的激光场被用来激发荧光团和三维成像的分子。当单个分子在多孔凝胶结构内移动并穿过多孔凝胶结构时，对其进行跟踪。与干燥的聚合物宿主相反，这些水基凝胶可以形成用于体外生物系统的单分子研究的有用介质。
Individual fluorescent molecules and individual singly labeled proteins were observed in the water-filled pores oi poly(acrylamide) gels by tar-field microscopy. Brownian motion was markedly reduced by the gel framework, thus enabling extended study of single fluorophores in aqueous environments. A highly axially dependent laser field was used both to excite the fluorophores and to image the molecules in three dimensions. Single molecules were followed as they moved within and through the porous gel structure. In contrast to dry polymeric hosts, these water-based gels may form a useful medium for single-molecule studies of biological systems in vitro.