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Title: Improved deconvolution of very weak confocal signals

Abstract

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 .

Authors:
; ; ; ; ; ORCiD logo
Publication Date:
Sponsoring Org.:
USDOE
OSTI Identifier:
1484822
Grant/Contract Number:  
AC02-06CH11357
Resource Type:
Journal Article: Published Article
Journal Name:
F1000Research
Additional Journal Information:
Journal Name: F1000Research Journal Volume: 6; Journal ID: ISSN 2046-1402
Publisher:
F1000 Research, Ltd.
Country of Publication:
United Kingdom
Language:
English

Citation Formats

Day, Kasey J., La Rivière, Patrick J., Chandler, Talon, Bindokas, Vytas P., Ferrier, Nicola J., and Glick, Benjamin S.. Improved deconvolution of very weak confocal signals. United Kingdom: N. p., 2017. Web. doi:10.12688/f1000research.11773.1.
Day, Kasey J., La Rivière, Patrick J., Chandler, Talon, Bindokas, Vytas P., Ferrier, Nicola J., & Glick, Benjamin S.. Improved deconvolution of very weak confocal signals. United Kingdom. doi:10.12688/f1000research.11773.1.
Day, Kasey J., La Rivière, Patrick J., Chandler, Talon, Bindokas, Vytas P., Ferrier, Nicola J., and Glick, Benjamin S.. Tue . "Improved deconvolution of very weak confocal signals". United Kingdom. doi:10.12688/f1000research.11773.1.
@article{osti_1484822,
title = {Improved deconvolution of very weak confocal signals},
author = {Day, Kasey J. and La Rivière, Patrick J. and Chandler, Talon and Bindokas, Vytas P. and Ferrier, Nicola J. and Glick, Benjamin S.},
abstractNote = {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 .},
doi = {10.12688/f1000research.11773.1},
journal = {F1000Research},
number = ,
volume = 6,
place = {United Kingdom},
year = {Tue Jun 06 00:00:00 EDT 2017},
month = {Tue Jun 06 00:00:00 EDT 2017}
}

Journal Article:
Free Publicly Available Full Text
Publisher's Version of Record at 10.12688/f1000research.11773.1

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Works referenced in this record:

Quantitative Comparison of Algorithms for Tracking Single Fluorescent Particles
journal, October 2001

  • Cheezum, Michael K.; Walker, William F.; Guilford, William H.
  • Biophysical Journal, Vol. 81, Issue 4, p. 2378-2388
  • DOI: 10.1016/S0006-3495(01)75884-5