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Journal of Biomedical Optics • Open Access

Integrated Raman and angular scattering microscopy reveals chemical and morphological differences between activated and nonactivated CD8+ T lymphocytes
Author(s): Zachary J. Smith; Jyh-Chiang E. Wang; Sally A. Quataert; Andrew J. Berger

Paper Abstract

Integrated Raman and angular-scattering microscopy (IRAM) is a multimodal platform capable of noninvasively probing both the chemistry and morphology of a single cell without prior labeling. Using this system, we are able to detect activation-dependent changes in the Raman and elastic-scattering signals from CD8+ T cells stimulated with either Staphylococcal enterotoxin B (SEB) or phorbol myristate acetate (PMA). In both cases, results obtained from the IRAM instrument correlate well with results obtained from traditional fluorescence-based flow cytometry for paired samples. SEB-mediated activation was distinguished from resting state in CD8+ T cells by an increase in the number and mean size of small (~500-nm) elastic scatterers as well as a decrease in Raman bands, indicating changes in nuclear content. PMA-mediated activation induced a different profile in CD8+ T cells from SEB, showing a similar increase in small elastic scatterers but a different Raman change, with elevation of cellular protein and lipid bands. These results suggest the potential of this multimodal, label-free optical technique for studying processes in single cells.

Paper Details

Date Published: 1 May 2010
PDF: 11 pages
J. Biomed. Opt. 15(3) 036021 doi: 10.1117/1.3443794
Published in: Journal of Biomedical Optics Volume 15, Issue 3
Show Author Affiliations
Zachary J. Smith, Univ. of Rochester (United States)
Jyh-Chiang E. Wang, Univ. of Rochester Medical Ctr. (United States)
Sally A. Quataert, Univ. of Rochester Medical Ctr. (United States)
Andrew J. Berger, Univ. of Rochester (United States)

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