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Noise‐Corrected Principal Component Analysis of fluorescence lifetime imaging data
Author(s) -
Le Marois Alix,
Labouesse Simon,
Suhling Klaus,
Heintzmann Rainer
Publication year - 2017
Publication title -
journal of biophotonics
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.877
H-Index - 66
eISSN - 1864-0648
pISSN - 1864-063X
DOI - 10.1002/jbio.201600160
Subject(s) - principal component analysis , noise (video) , fluorescence lifetime imaging microscopy , membrane , hela , photon counting , biological system , chemistry , fluorescence , analytical chemistry (journal) , photon , physics , optics , cell , artificial intelligence , biology , computer science , chromatography , biochemistry , image (mathematics)
Fluorescence Lifetime Imaging (FLIM) is an attractive microscopy method in the life sciences, yielding information on the sample otherwise unavailable through intensity‐based techniques. A novel Noise‐Corrected Principal Component Analysis (NC‐PCA) method for time‐domain FLIM data is presented here. The presence and distribution of distinct microenvironments are identified at lower photon counts than previously reported, without requiring prior knowledge of their number or of the dye's decay kinetics. A noise correction based on the Poisson statistics inherent to Time‐Correlated Single Photon Counting is incorporated. The approach is validated using simulated data, and further applied to experimental FLIM data of HeLa cells stained with membrane dye di‐4‐ANEPPDHQ. Two distinct lipid phases were resolved in the cell membranes, and the modification of the order parameters of the plasma membrane during cholesterol depletion was also detected.Noise‐corrected Principal Component Analysis of FLIM data resolves distinct microenvironments in cell membranes of live HeLa cells.