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Biomedical Optics Express

Biomedical Optics Express

  • Editor: Joseph A. Izatt
  • Vol. 5, Iss. 5 — May. 1, 2014
  • pp: 1378–1390

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

Claudia Di Napoli, Iestyn Pope, Francesco Masia, Peter Watson, Wolfgang Langbein, and Paola Borri  »View Author Affiliations

Biomedical Optics Express, Vol. 5, Issue 5, pp. 1378-1390 (2014)

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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.

© 2014 Optical Society of America

OCIS Codes
(300.6230) Spectroscopy : Spectroscopy, coherent anti-Stokes Raman scattering
(180.5655) Microscopy : Raman microscopy

ToC Category:

Original Manuscript: February 11, 2014
Revised Manuscript: March 9, 2014
Manuscript Accepted: March 9, 2014
Published: April 2, 2014

Claudia Di Napoli, Iestyn Pope, Francesco Masia, Peter Watson, Wolfgang Langbein, and Paola Borri, "Hyperspectral and differential CARS microscopy for quantitative chemical imaging in human adipocytes," Biomed. Opt. Express 5, 1378-1390 (2014)

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