Quantitative Vibrational Imaging by Hyperspectral Stimulated Raman Scattering Microscopy and Multivariate Curve Resolution Analysis

Anal. Chem., 2013, 85 (1), pp 98–106.

Delong Zhang , Ping Wang , Mikhail N. Slipchenko, Dor Ben-Amotz , Andrew M. Weiner , Ji-Xin Cheng

 

Department of Chemistry, Weldon School of Biomedical Engineering, and Electrical and Computer Engineering, Purdue University, West Lafayette, Indiana 47907, United States.

 

 

Abstract

Spectroscopic imaging has been an increasingly critical approach for unveiling specific molecules in biological environments. Toward this goal, we demonstrate hyperspectral stimulated Raman loss (SRL) imaging by intrapulse spectral scanning through a femtosecond pulse shaper. The hyperspectral stack of SRL images is further analyzed by a multivariate curve resolution (MCR) method to reconstruct quantitative concentration images for each individual component and retrieve the corresponding vibrational Raman spectra. Using these methods, we demonstrate quantitative mapping of dimethyl sulfoxide concentration in aqueous solutions and in fat tissue. Moreover, MCR is performed on SRL images of breast cancer cells to generate maps of principal chemical components along with their respective vibrational spectra. These results show the great capability and potential of hyperspectral SRL microscopy for quantitative imaging of complicated biomolecule mixtures through resolving overlapped Raman bands.

Copyright © 2012 American Chemical Society

 

 

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Figure Legend

Label-free Imaging of Tumor cells by Hyperspectral Stimulated Raman Scattering Microscopy and Multivariate Curve Resolution Analysis.

 

Quantitative Vibrational Imaging by Hyperspectral Stimulated Raman Scattering Microscopy and Multivariate Curve Resolution Analysis

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