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

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...

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Bibliographic Details
Published in:Analytical chemistry (Washington) 2013-01, Vol.85 (1), p.98-106
Main Authors: Zhang, Delong, Wang, Ping, Slipchenko, Mikhail N, Ben-Amotz, Dor, Weiner, Andrew M, Cheng, Ji-Xin
Format: Article
Language:English
Subjects:
Adipose Tissue - chemistry
Analytical chemistry
Animals
Aqueous solutions
Breast cancer
Cells
Dimethyl Sulfoxide - analysis
Humans
MCF-7 Cells
Mice
Mice, Inbred C57BL
Molecules
Multivariate Analysis
Principal Component Analysis
Spectrum Analysis, Raman
Vibration
Water - chemistry
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Summary: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.
ISSN:0003-2700
1520-6882
DOI:10.1021/ac3019119