Mechanical stability of a microscope setup working at a few kelvins for single-molecule localization
Mechanical stability of a microscope setup working at a few kelvins for single-molecule localization
复制标题
DOI：
10.1016/j.chemphys.2013.02.024
复制
发表时间：
2013-06-20
期刊：
影响因子：
2.3
通讯作者：
Fujiyoshi, Satoru
中科院分区：
文献类型：
--
作者：
Hinohara, Takuya;Hamada, Yuki I.;Fujiyoshi, Satoru
单分子荧光成像的一个巨大优势是分子的定位精度超过衍射极限。虽然较长的信号采集时间可以获得较高的精度，但室温下的采集时间通常受到光漂白、热扩散等因素的限制。在几K的低温下，如果样品和物镜保持足够稳定，则可以获得更长的采集时间，并将提高精度。本工作研究了保持稳定的样品和目标在1.5 K的超流氦在氦浴。通过聚合物珠的点散射源的定位精度来评价稳定性。散射光由物镜收集，并由自制的刚性成像单元成像。对于在17分钟内连续拍摄的800个图像确定的质心位置的标准偏差在水平方向上为0.5nm，在垂直方向上为0.9nm。(C)2013年爱思唯尔B。V.保留所有权利。
A great advantage of single-molecule fluorescence imaging is the localization precision of molecule beyond the diffraction limit. Although longer signal-acquisition yields higher precision, acquisition time at room temperature is normally limited by photobleaching, thermal diffusion, and so on. At low temperature of a few kelvins, much longer acquisition is possible and will improve precision if the sample and the objective are held stably enough. The present work examined holding stability of the sample and objective at 1.5 K in superfluid helium in the helium bath. The stability was evaluated by localization precision of a point scattering source of a polymer bead. Scattered light was collected by the objective, and imaged by a home-built rigid imaging unit. The standard deviation of the centroid position determined for 800 images taken continuously in 17 min was 0.5 nm in the horizontal and 0.9 nm in the vertical directions. (C) 2013 Elsevier B. V. All rights reserved.