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Proceedings Paper

Raman spectra of single cells with autofluorescence suppression by modulated wavelength excitation
Author(s): Christoph Krafft; Sebastian Dochow; Norbert Bergner; Joachim H. Clement; Bavishna B. Praveen; Michael Mazilu; Rob Marchington; Kishan Dholakia; Jürgen Popp
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Paper Abstract

Raman spectroscopy is a non-invasive technique offering great potential in the biomedical field for label-free discrimination between normal and tumor cells based on their biochemical composition. First, this contribution describes Raman spectra of lymphocytes after drying, in laser tweezers, and trapped in a microfluidic environment. Second, spectral differences between lymphocytes and acute myeloid leukemia cells (OCI-AML3) are compared for these three experimental conditions. Significant similarities of difference spectra are consistent with the biological relevance of the spectral features. Third, modulated wavelength Raman spectroscopy has been applied to this model system to demonstrate background suppression. Here, the laser excitation wavelength of 785 nm was modulated with a frequency of 40 mHz by 0.6 nm. 40 spectra were accumulated with an exposure time of 5 seconds each. These data were subjected to principal component analysis to calculate modulated Raman signatures. The loading of the principal component shows characteristics of first derivatives with derivative like band shapes. The derivative of this loading corresponds to a pseudo-second derivative spectrum and enables to determine band positions.

Paper Details

Date Published: 31 January 2012
PDF: 7 pages
Proc. SPIE 8219, Biomedical Vibrational Spectroscopy V: Advances in Research and Industry, 82190F (31 January 2012); doi: 10.1117/12.908564
Show Author Affiliations
Christoph Krafft, Institute of Photonic Technology (Germany)
Sebastian Dochow, Institute of Photonic Technology (Germany)
Norbert Bergner, Institute of Photonic Technology (Germany)
Joachim H. Clement, Univ. Hospital Jena (Germany)
Bavishna B. Praveen, Univ. of St. Andrews (United States)
Michael Mazilu, Univ. of St. Andrews (United Kingdom)
Rob Marchington, Univ. of St. Andrews (United Kingdom)
Kishan Dholakia, Univ. of St. Andrews (United Kingdom)
Jürgen Popp, Institute of Photonic Technology (Germany)
Univ. Jena (Germany)

Published in SPIE Proceedings Vol. 8219:
Biomedical Vibrational Spectroscopy V: Advances in Research and Industry
Anita Mahadevan-Jansen; Wolfgang Petrich, Editor(s)

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