Hyperspectral and differential CARS microscopy for quantitative chemical imaging in human adipocytes

Di Napoli, Claudia, Pope, Iestyn ORCID: https://orcid.org/0000-0002-4104-0389, Masia, Francesco ORCID: https://orcid.org/0000-0003-4958-410X, Watson, Peter Duncan ORCID: https://orcid.org/0000-0003-0250-7852, Langbein, Wolfgang Werner ORCID: https://orcid.org/0000-0001-9786-1023 and Borri, Paola ORCID: https://orcid.org/0000-0002-7873-3314 2014. Hyperspectral and differential CARS microscopy for quantitative chemical imaging in human adipocytes. Biomedical Optics Express 5 (5) , pp. 1378-1390. 10.1364/BOE.5.001378

Preview

PDF - Published Version Download (2MB) | Preview

Abstract

In this work, we demonstrate the applicability of coherent anti-Stokes Raman scattering (CARS) micro-spectroscopy for quantitative chemical imaging of saturated and unsaturated lipids in human stem-cell derived adipocytes. We compare dual-frequency/differential CARS (D-CARS), which enables rapid imaging and simple data analysis, with broadband hyperspectral CARS microscopy analyzed using an unsupervised phase-retrieval and factorization method recently developed by us for quantitative chemical image analysis. Measurements were taken in the vibrational fingerprint region (1200–2000/cm) and in the CH stretch region (2600–3300/cm) using a home-built CARS set-up which enables hyperspectral imaging with 10/cm resolution via spectral focussing from a single broadband 5 fs Ti:Sa laser source. Through a ratiometric analysis, both D-CARS and phase-retrieved hyperspectral CARS determine the concentration of unsaturated lipids with comparable accuracy in the fingerprint region, while in the CH stretch region D-CARS provides only a qualitative contrast owing to its non-linear behavior. When analyzing hyperspectral CARS images using the blind factorization into susceptibilities and concentrations of chemical components recently demonstrated by us, we are able to determine vol:vol concentrations of different lipid components and spatially resolve inhomogeneities in lipid composition with superior accuracy compared to state-of-the art ratiometric methods.

Item Type:	Article
Date Type:	Publication
Status:	Published
Schools:	Physics and Astronomy Biosciences
Publisher:	OSA
ISSN:	2156-7085
Date of First Compliant Deposit:	30 March 2016
Date of Acceptance:	9 March 2014
Last Modified:	15 Sep 2023 16:25
URI:	https://orca.cardiff.ac.uk/id/eprint/58703

Citation Data

Cited 46 times in Scopus. View in Scopus. Powered By Scopus® Data

Actions (repository staff only)

Edit Item