Hyperspectral imaging and characterization of live cells by broadband coherent anti-stokes raman scattering (CARS) microscopy with singular value decomposition (SVD) analysis

Alexander Khmaladze, Joshua Jasensky, Erika Price, Chi Zhang, Andrew Boughton, Xiaofeng Han, Emily Seeley, Xinran Liu, Mark M.Banaszak Holl, Zhan Chen

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Abstract

Coherent anti-Stokes Raman scattering (CARS) microscopy can be used as a powerful imaging technique to identify chemical compositions of complex samples in biology, biophysics, medicine, and materials science. In this work we developed a CARS microscopic system capable of hyperspectral imaging. By employing an ultrafast laser source, a photonic crystal fiber, and a scanning laser microscope together with spectral detection by a highly sensitive back-illuminated cooled charge-coupled device (CCD) camera, we were able to rapidly acquire and process hyperspectral images of live cells with chemical selectivity. We discuss various aspects of hyperspectral CARS image analysis and demonstrate the use of singular value decomposition methods to characterize the cellular lipid content.

Original languageEnglish
Pages (from-to)1116-1122
Number of pages7
JournalApplied Spectroscopy
Volume68
Issue number10
DOIs
Publication statusPublished - 1 Jan 2014
Externally publishedYes

Keywords

  • Coherent anti-Stokes Raman scattering spectroscopy
  • Imaging systems
  • Raman microscopy
  • Scanning microscopy

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