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Proc Natl Acad Sci U S A. 2013 Oct 22;110(43):17344-9. doi: 10.1073/pnas.1315675110. Epub 2013 Oct 8.

High-resolution restoration of 3D structures from widefield images with extreme low signal-to-noise-ratio.

Author information

1
Department of Biochemistry and Biophysics and Keck Advanced Microscopy Center, University of California, San Francisco, CA 94158.

Abstract

Four-dimensional fluorescence microscopy--which records 3D image information as a function of time--provides an unbiased way of tracking dynamic behavior of subcellular components in living samples and capturing key events in complex macromolecular processes. Unfortunately, the combination of phototoxicity and photobleaching can severely limit the density or duration of sampling, thereby limiting the biological information that can be obtained. Although widefield microscopy provides a very light-efficient way of imaging, obtaining high-quality reconstructions requires deconvolution to remove optical aberrations. Unfortunately, most deconvolution methods perform very poorly at low signal-to-noise ratios, thereby requiring moderate photon doses to obtain acceptable resolution. We present a unique deconvolution method that combines an entropy-based regularization function with kernels that can exploit general spatial characteristics of the fluorescence image to push the required dose to extreme low levels, resulting in an enabling technology for high-resolution in vivo biological imaging.

KEYWORDS:

4D microscopy; low dose microscopy; noise-suppressing regularization

PMID:
24106307
PMCID:
PMC3808589
DOI:
10.1073/pnas.1315675110
[Indexed for MEDLINE]
Free PMC Article

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