4 March 2014 Enhanced 3D localization of individual RNA transcripts via astigmatic imaging
Author Affiliations +
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.
© (2014) COPYRIGHT Society of Photo-Optical Instrumentation Engineers (SPIE). Downloading of the abstract is permitted for personal use only.
Evan P. Perillo, Leyma De Haro, Mary E. Phipps, Jennifer S. Martinez, Hsin-Chih Yeh, Andrew K. Dunn, Douglas P. Shepherd, James H. Werner, "Enhanced 3D localization of individual RNA transcripts via astigmatic imaging", Proc. SPIE 8950, Single Molecule Spectroscopy and Superresolution Imaging VII, 895003 (4 March 2014); doi: 10.1117/12.2038197; https://doi.org/10.1117/12.2038197


New waves in light microscopy
Proceedings of SPIE (August 17 1994)
Advances in three-dimensional SAR from practical apertures
Proceedings of SPIE (November 05 1996)
Three-dimensional digital breast histopathology imaging
Proceedings of SPIE (April 12 2005)
Resolution in 3D in multifocal plane microscopy
Proceedings of SPIE (February 12 2008)

Back to Top