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Chemically‐specific dual/differential CARS micro‐spectroscopy of saturated and unsaturated lipid droplets
Author(s) -
Di Napoli Claudia,
Masia Francesco,
Pope Iestyn,
Otto Cees,
Langbein Wolfgang,
Borri Paola
Publication year - 2014
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.201200197
Subject(s) - raman spectroscopy , chemistry , spectroscopy , analytical chemistry (journal) , femtosecond , materials science , chemical engineering , chromatography , optics , laser , physics , quantum mechanics , engineering
We have investigated the ability of dual‐frequency Coherent Antistokes Raman Scattering (D‐CARS) micro‐spectroscopy, based on femtosecond pulses (100 fs or 5 fs) spectrally focussed by glass dispersion, to distinguish the chemical composition of micron‐sized lipid droplets consisting of different triglycerides types (poly‐unsaturated glyceryl trilinolenate, mono‐unsaturated glyceryl trioleate and saturated glyceryl tricaprylate and glyceryl tristearate) in a rapid and label‐free way. A systematic comparison of Raman spectra with CARS and D‐CARS spectra was used to identify D‐CARS spectral signatures which distinguish the disordered poly‐unsaturated lipids from the more ordered saturated ones both in the CH‐stretch vibration region and in the fingerprint region, without the need for lengthy CARS multiplex acquisition and analysis. D‐CARS images of the lipid droplets at few selected wavenumbers clearly resolved the lipid composition differences, and exemplify the potential of this technique for label‐free chemically selective rapid imaging of cytosolic lipid droplets in living cells. (© 2014 WILEY‐VCH Verlag GmbH & Co. KGaA, Weinheim)