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Proceedings Paper

Double helix PSF engineering for computational fluorescence microscopy imaging
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Paper Abstract

Point spread function engineering with a double helix (DH) phase mask has been recently used in a joint computationaloptical approach for the determination of depth and intensity information from fluorescence images. In this study, theoretically determined DH-PSFs computed from a model that incorporates different amounts of depth-induced spherical aberration (SA) due to refractive-index mismatch in the three-dimensional imaging layers, are evaluated through a comparison to empirically determined DH-PSFs measured from quantum dots. The theoretically-determined DH-PSFs show a trend that captures the main effects observed in the empirically-determined DH-PSFs. Calibration curves computed from these DH-PSFs show that SA slows down the rate of rotation observed in a DH-PSF which results in: 1) an extended range of rotation; and 2) asymmetric rotation ranges as the focus is moved in opposite directions. Thus, for accurate particle localization different calibration curves need to be known for different amounts of SA. Results also show that the DH-PSF is less sensitive to SA than the conventional PSF. Based on this result, it is expected that fewer depth-variant (DV) DH-PSFs will be required for 3D computational microscopy imaging in the presence of SA compared to the required number of conventional DV PSFs.

Paper Details

Date Published: 2 February 2012
PDF: 7 pages
Proc. SPIE 8227, Three-Dimensional and Multidimensional Microscopy: Image Acquisition and Processing XIX, 82270F (2 February 2012); doi: 10.1117/12.909757
Show Author Affiliations
Sreya Ghosh, The Univ. of Memphis (United States)
Sean A. Quirin, Univ. of Colorado at Boulder (United States)
Ginni Grover, Univ. of Colorado at Boulder (United States)
Rafael Piestun, Univ. of Colorado at Boulder (United States)
Chrysanthe Preza, The Univ. of Memphis (United States)

Published in SPIE Proceedings Vol. 8227:
Three-Dimensional and Multidimensional Microscopy: Image Acquisition and Processing XIX
Jose-Angel Conchello; Carol J. Cogswell; Tony Wilson; Thomas G. Brown, Editor(s)

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