Nanometer Accuracy in Cryogenic Far-Field Localization Microscopy of Individual Molecules
Nanometer Accuracy in Cryogenic Far-Field Localization Microscopy of Individual Molecules
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单个分子低温远场定位显微镜的纳米精度
DOI：
10.1021/acs.jpclett.9b02184
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发表时间：
2019
期刊：
影响因子：
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通讯作者：
Fujiyoshi Satoru
中科院分区：
文献类型：
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作者：
Furubayashi Taku;Ishida Keita;Kashida Hiromu;Nakata Eiji;Morii Takashi;Matsushita Michio;Fujiyoshi Satoru
我们利用近红外和红色荧光团结合到双链DNA分子（10.2 nm长度），在1.8 K温度下证明了远场荧光定位显微镜的纳米精度。虽然每个荧光团在±0.7 μm的焦深度内以1 nm的横向精度定位，但两个荧光团在横向平面上的距离（Dxy）分布在0到50 nm之间。这种系统误差主要是由于用大焦深探测定向固定荧光团的偶极子发射引起的。每个荧光团定位精度为±1 nm（横向），同时±11 nm（轴向），在轴向（- 900至900 nm）每100 nm采集图像。通过修正偶极子取向效应，dxy的分布以DNA长度为中心。dxy的平均值为10 nm，标准差为5 nm。
We demonstrate the nanometer accuracy of far-field fluorescence localization microscopy at a temperature of 1.8 K using near-infrared and red fluorophores bonded to double-stranded DNA molecules (10.2 nm length). Although each fluorophore was localized with a 1 nm lateral precision by acquiring an image at one axial position within the focal depth of ±0.7 μm, the distance between the two fluorophores on the lateral plane (Dxy) was distributed from 0 to 50 nm. This systematic error was mainly due to detecting with the large focal depth the dipole emission from orientationally fixed fluorophores. Each fluorophore was localized with precisions of ±1 nm (lateral) and simultaneously ±11 nm (axial) by acquiring images every 100 nm in the axial direction from −900 to 900 nm. By correcting the dipole orientation effects, the distribution ofDxywas centered around the DNA length. The average and standard deviation ofDxywere 10 and 5 nm.