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Proceedings Paper

Enhanced 3D localization of individual RNA transcripts via astigmatic imaging
Author(s): Evan P. Perillo; Leyma De Haro; Mary E. Phipps; Jennifer S. Martinez; Hsin-Chih Yeh; Andrew K. Dunn; Douglas P. Shepherd; James H. Werner
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Paper Abstract

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.

Paper Details

Date Published: 4 March 2014
PDF: 11 pages
Proc. SPIE 8950, Single Molecule Spectroscopy and Superresolution Imaging VII, 895003 (4 March 2014); doi: 10.1117/12.2038197
Show Author Affiliations
Evan P. Perillo, The Univ. of Texas at Austin (United States)
Leyma De Haro, Los Alamos National Lab. (United States)
Mary E. Phipps, Los Alamos National Lab. (United States)
Jennifer S. Martinez, Los Alamos National Lab. (United States)
Hsin-Chih Yeh, The Univ. of Texas at Austin (United States)
Andrew K. Dunn, The Univ. of Texas at Austin (United States)
Douglas P. Shepherd, Los Alamos National Lab. (United States)
Univ. of Colorado, Denver (United States)
James H. Werner, Los Alamos National Lab. (United States)


Published in SPIE Proceedings Vol. 8950:
Single Molecule Spectroscopy and Superresolution Imaging VII
Jörg Enderlein; Ingo Gregor; Zygmunt Karol Gryczynski; Rainer Erdmann; Felix Koberling, Editor(s)

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