Using spectral decomposition of the signals from laurdan-derived probes to evaluate the physical state of membranes in live cells.

ABSTRACT: Background: We wanted to investigate the physical state of biological membranes in live cells under the most physiological conditions possible. Methods: For this we have been using laurdan, C-laurdan or M-laurdan to label a variety of cells, and a biphoton microscope equipped with both a thermostatic chamber and a spectral analyser. We also used a flow cytometer to quantify the 450/530 nm ratio of fluorescence emissions by whole cells. Results: We find that using all the information provided by spectral analysis to perform spectral decomposition dramatically improves the imaging resolution compared to using just two channels, as commonly used to calculate generalized polarisation (GP). Coupled to a new plugin called Fraction Mapper, developed to represent the fraction

SUBMITTER: Mazeres S 

PROVIDER: S-EPMC5473435 | biostudies-literature | 2017

REPOSITORIES: biostudies-literature