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Sci Rep. 2017 Aug 23;7(1):9215. doi: 10.1038/s41598-017-08564-z.

A multidimensional phasor approach reveals LAURDAN photophysics in NIH-3T3 cell membranes.

Author information

1
Laboratory for Fluorescence Dynamics, Biomedical Engineering Department, University of California at Irvine, Irvine, California, USA.
2
Área de Investigación Respiratoria, Departamento de Fisiopatología, Hospital de Clínicas, Facultad de Medicina, Universidad de la República, Montevideo, Uruguay.
3
Analytical Biochemistry and Proteomics Unit, Institut Pasteur of Montevideo, Montevideo, Uruguay.
4
Department Cell and Molecular Biology, University of Hawai'i at Manoa, John A. Burns School of Medicine, Honolulu, HI, USA.
5
Laboratory for Fluorescence Dynamics, Biomedical Engineering Department, University of California at Irvine, Irvine, California, USA. egratton22@gmail.com.

Abstract

Mammalian cell membranes have different phospholipid composition and cholesterol content, displaying a profile of fluidity that depends on their intracellular location. Among the dyes used in membrane studies, LAURDAN has the advantage to be sensitive to the lipid composition as well as to membrane fluidity. The LAURDAN spectrum is sensitive to the lipid composition and dipolar relaxation arising from water penetration, but disentangling lipid composition from membrane fluidity can be obtained if time resolved spectra could be measured at each cell location. Here we describe a method in which spectral and lifetime information obtained in different measurements at the same plane in a cell are used in the phasor plot providing a solution to analyze multiple lifetime or spectral data through a common visualization approach. We exploit a property of phasor plots based on the reciprocal role of the phasor plot and the image. In the phasor analysis each pixel of the image is associated with a phasor and each phasor maps to pixels and features in the image. In this paper the lifetime and spectral fluorescence data are used simultaneously to determine the contribution of polarity and dipolar relaxations of LAURDAN in each pixel of an image.

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