title: Quantitative vibrational imaging by hyperspectral stimulated Raman scattering microscopy and multivariate curve resolution analysis.
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.
authors.affiliation: Department of Chemistry, Purdue University, West Lafayette, Indiana 47907, United States.
authors.forename: Delong
authors.initials: D
authors.lastname: Zhang
authors.forename: Ping
authors.initials: P
authors.lastname: Wang
authors.forename: Mikhail N
authors.initials: MN
authors.lastname: Slipchenko
authors.forename: Dor
authors.initials: D
authors.lastname: Ben-Amotz
authors.forename: Andrew M
authors.initials: AM
authors.lastname: Weiner
authors.forename: Ji-Xin
authors.initials: JX
authors.lastname: Cheng
mesh_terms: D000273:Adipose Tissue / Q000737:chemistry; D000818:Animals; D004121:Dimethyl Sulfoxide / Q000032:analysis; D006801:Humans; D061986:MCF-7 Cells; D051379:Mice; D008810:Mice, Inbred C57BL; D015999:Multivariate Analysis; D025341:Principal Component Analysis; D013059:Spectrum Analysis, Raman; D014732:Vibration; D014867:Water / Q000737:chemistry
chemical_list: D014867:Water; D004121:Dimethyl Sulfoxide