Single-photon atomic force microscopy

Single-photon atomic force microscopy
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单光子原子力显微镜

DOI:
10.1007/s00216-009-3426-0
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发表时间:
2010-01
期刊:
Anal Bioanal Chem
影响因子:
--
通讯作者:
张军
张军
中科院分区:
其他
文献类型:
--
作者:
张军

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在过去的几年中,一系列新技术,特别是扫描探针显微镜大家族,现在经常与其他强大的成像工具(例如激光共焦显微镜和全内反射荧光显微镜)集成,已广泛应用于生物分子相互作用和动力学的研究。但以高时空分辨率直接监测活细胞中生物分子相互作用的动态仍然是一个巨大的挑战。在原子力显微镜与碳纳米管和单光子检测技术相结合的基础上,提出了一种称为“单光子原子力显微镜”(SP-AFM)的创新方法,该方法优于现有技术来追踪体内生物分子相互作用和动力学。 SP-AFM作为一种独特的工具,能够在扫描样品的同时同步采集和分析表面形貌和荧光光信号,从而实现亚纳米级的同步形貌成像和分子识别,在探索活细胞或复杂生物分子背景中的生物分子相互作用和动力学方面可以发挥非常重要的作用。图示意性地显示了SP-AFM同步分子成像和识别的过程。 (A) 三个球代表扫描区域中生物样品(例如膜等)表面上具有内源性或选择性标记的外源性荧光团的不同蛋白质。在样品扫描的同时,SP-AFM可以同步获取表面形貌数据(B)和荧光光信号(C)。通过数据处理,SP-AFM 能够在一次扫描中同时成像和识别三种分子 (D)。
In the last few years, an array of novel technologies, especially the big family of scanning probe microscopy, now often integrated with other powerful imaging tools such as laser confocal microscopy and total internal reflection fluorescence microscopy, have been widely applied in the investigation of biomolecular interactions and dynamics. But it is still a great challenge to directly monitor the dynamics of biomolecular interactions with high spatial and temporal resolution in living cells. An innovative method termed “single-photon atomic force microscopy” (SP-AFM), superior to existing techniques in tracing biomolecular interactions and dynamics in vivo, was proposed on the basis of the combination of atomic force microscopy with the technologies of carbon nanotubes and single-photon detection. As a unique tool, SP-AFM, capable of simultaneous topography imaging and molecular identification at the subnanometer level by synchronous acquisitions and analyses of the surface topography and fluorescent optical signals while scanning the sample, could play a very important role in exploring biomolecular interactions and dynamics in living cells or in a complicated biomolecular background.FigureThe figure schematically showed the process of simultaneous molecular imaging and identification with SP-AFM. (A) The three balls represented different proteins with endogenic or selectively labelled exogenous fluorophores on the surface of a biological sample such as membrane, etc. in a scanning area. While sample scanning, the SP-AFM could synchronously acquire the data of the surface topography (B) and fluorescent optical signals (C). By data processing, the SP-AFM was capable of simultaneous imaging and identification of the three molecules in one scanning (D).
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