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Image reconstruction enables high resolution imaging at large penetration depths in fluorescence microscopy

Dilipkumar, Shilpa and Montalescot, Sandra and Mondal, Partha Pratim (2013) Image reconstruction enables high resolution imaging at large penetration depths in fluorescence microscopy. In: APPLIED PHYSICS LETTERS, 103 (17).

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Official URL: http://dx.doi.org/10.1063/1.4827191


Imaging thick specimen at a large penetration depth is a challenge in biophysics and material science. Refractive index mismatch results in spherical aberration that is responsible for streaking artifacts, while Poissonian nature of photon emission and scattering introduces noise in the acquired three-dimensional image. To overcome these unwanted artifacts, we introduced a two-fold approach: first, point-spread function modeling with correction for spherical aberration and second, employing maximum-likelihood reconstruction technique to eliminate noise. Experimental results on fluorescent nano-beads and fluorescently coated yeast cells (encaged in Agarose gel) shows substantial minimization of artifacts. The noise is substantially suppressed, whereas the side-lobes (generated by streaking effect) drops by 48.6% as compared to raw data at a depth of 150 mu m. Proposed imaging technique can be integrated to sophisticated fluorescence imaging techniques for rendering high resolution beyond 150 mu m mark. (C) 2013 AIP Publishing LLC.

Item Type: Journal Article
Additional Information: Copyright for this article belongs to the AMER INST PHYSICS USA
Department/Centre: Division of Physical & Mathematical Sciences > Physics
Date Deposited: 26 Dec 2013 05:17
Last Modified: 26 Dec 2013 05:17
URI: http://eprints.iisc.ac.in/id/eprint/47974

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