Enhanced 3D localization of individual RNA transcripts via astigmatic imaging

被引：0
作者：
Perillo, Evan P. [1 ]
De Haro, Leyma [2 ]
Phipps, Mary E. [2 ]
Martinez, Jennifer S. [2 ]
Yeh, Hsin-Chih [1 ]
Dunn, Andrew K. [1 ]
Shepherd, Douglas P. [2 ,3 ]
Werner, James H. [2 ]
机构：
[1] Univ Texas Austin, Dept Biomed Engn, Austin, TX 78712 USA
[2] Los Alamos Natl Lab, Ctr Intergrated Nanotechnol, Los Alamos, NM 87544 USA
[3] Univ Colorado, Dept Phys, Denver, CO 80204 USA
来源：
SINGLE MOLECULE SPECTROSCOPY AND SUPERRESOLUTION IMAGING VII | 2014年 / 8950卷
关键词：
smFISH; single-molecule localization; astigmatic imaging; fluorescence microscopy; gene expression; GENE-EXPRESSION; STEM-CELLS; SINGLE; OSTEOBLAST; REVEALS; DIFFERENTIATION;
D O I：
10.1117/12.2038197
中图分类号：
TH7 [仪器、仪表];
学科分类号：
0804 ; 080401 ; 081102 ;
摘要：
Here we present an automated microscope capable of 3D multi-color single molecule localization of individual messenger RNA molecules in a wide range of cell types. We have implemented astigmatic imaging with a cylindrical lens to improve z-localization, and a maximum likelihood estimator on a graphics processing unit to improve localization precision and speed. This microscope will aid in gene expression analysis by its capability to perform high throughput imaging of thick cells and tissues while still maintaining sufficient z resolution to resolve single RNA transcripts in three dimensions. Enhanced z-localization allows for resolving membrane localized and co-localized transcripts.
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页数：11
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