1 May 2010 Integrated Raman and angular scattering microscopy reveals chemical and morphological differences between activated and nonactivated CD8+ T lymphocytes
Author Affiliations +
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.
© (2010) Society of Photo-Optical Instrumentation Engineers (SPIE)
Zachary J. Smith, Zachary J. Smith, Jyh-Chiang E. Wang, Jyh-Chiang E. Wang, Sally A. Quataert, Sally A. Quataert, Andrew J. Berger, Andrew J. Berger, } "Integrated Raman and angular scattering microscopy reveals chemical and morphological differences between activated and nonactivated CD8+ T lymphocytes," Journal of Biomedical Optics 15(3), 036021 (1 May 2010). https://doi.org/10.1117/1.3443794 . Submission:
JOURNAL ARTICLE
11 PAGES


SHARE
RELATED CONTENT

Label-free high-throughput imaging flow cytometry
Proceedings of SPIE (March 06 2014)
Structural Raman enhancement in graphite nano-discs
Proceedings of SPIE (April 17 2016)
CARS microscope enables BISTRO measurements
Proceedings of SPIE (February 20 2017)
Integrated Raman and angular scattering microscopy (IRAM)
Proceedings of SPIE (March 09 2008)

Back to Top