Journal
JOURNAL OF BIOPHOTONICS
Volume 5, Issue 5-6, Pages 449-457Publisher
WILEY-V C H VERLAG GMBH
DOI: 10.1002/jbio.201100137
Keywords
Time-gated fluorescence lifetime imaging microscopy; precision improvement; image denoising; wavelet analysis; total variation
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Funding
- National Institutes of Health [R01-CA-114542]
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We report the first application of wavelet-based denoising (noise removal) methods to time-domain box-car fluorescence lifetime imaging microscopy (FLIM) images and compare the results to novel total variation (TV) denoising methods. Methods were tested first on artificial images and then applied to low-light live-cell images. Relative to undenoised images, TV methods could improve lifetime precision up to 10-fold in artificial images, while preserving the overall accuracy of lifetime and amplitude values of a single-exponential decay model and improving local lifetime fitting in live-cell images. Wavelet-based methods were at least 4-fold faster than TV methods, but could introduce significant inaccuracies in recovered lifetime values. The denoising methods discussed can potentially enhance a variety of FLIM applications, including live-cell, in vivo animal, or endoscopic imaging studies, especially under challenging imaging conditions such as low-light or fast video-rate imaging. (C) 2012 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim)
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