Improved deconvolution of very weak confocal signals

Day, Kasey J.; La Rivière, Patrick J.; Chandler, Talon; Bindokas, Vytas P.; Ferrier, Nicola J.; Glick, Benjamin S.

doi:10.12688/f1000research.11773.2

Improved deconvolution of very weak confocal signals

Journal Article · Mon Aug 07 00:00:00 EDT 2017 · F1000Research

DOI:https://doi.org/10.12688/f1000research.11773.2· OSTI ID:1484821

Day, Kasey J.; La Rivière, Patrick J.; Chandler, Talon; Bindokas, Vytas P.; Ferrier, Nicola J.;

Deconvolution is typically used to sharpen fluorescence images, but when the signal-to-noise ratio is low, the primary benefit is reduced noise and a smoother appearance of the fluorescent structures. 3D time-lapse (4D) confocal image sets can be improved by deconvolution. However, when the confocal signals are very weak, the popular Huygens deconvolution software erases fluorescent structures that are clearly visible in the raw data. We find that this problem can be avoided by prefiltering the optical sections with a Gaussian blur. Analysis of real and simulated data indicates that the Gaussian blur prefilter preserves meaningful signals while enabling removal of background noise. This approach is very simple, and it allows Huygens to be used with 4D imaging conditions that minimize photodamage.

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Research Organization:: Argonne National Lab. (ANL), Argonne, IL (United States)

Sponsoring Organization:: USDOE; USDOE Office of Science (SC), Biological and Environmental Research (BER) (SC-23)

Grant/Contract Number:: AC02-06CH11357

OSTI ID:: 1484821

Alternate ID(s):: OSTI ID: 1395871

Journal Information:: F1000Research, Journal Name: F1000Research Vol. 6; ISSN 2046-1402

Publisher:: F1000 Research, Ltd.Copyright Statement

Country of Publication:: United Kingdom

Language:: English

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TIFF files for the experimental and simulated image data are provided in the compressed folder Original Image Files.zip. Day, Kasey J.; Rivière, Patrick J. La; Chandler, Talon F1000Research https://doi.org/10.5256/f1000research.11773.d163336	dataset	January 2017
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