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Journal of Biomedical Optics

Kinetics of T-cell receptor-dependent antigen recognition determined in vivo by multi-spectral normalized epifluorescence laser scanning
Author(s): Rosy Favicchio; Katerina Oikonomaki; Kleo Mamalaki; Giannis Zacharakis; Athanasios Zacharopoulos; Jorge Ripoll
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Paper Abstract

Detection of multiple fluorophores in conditions of low signal represents a limiting factor for the application of in vivo optical imaging techniques in immunology where fluorescent labels report for different functional characteristics. A noninvasive in vivo Multi-Spectral Normalized Epifluorescence Laser scanning (M-SNELS) method was developed for the simultaneous and quantitative detection of multiple fluorophores in low signal to noise ratios and used to follow T-cell activation and clonal expansion. Colocalized DsRed- and GFP-labeled T cells were followed in tandem during the mounting of an immune response. Spectral unmixing was used to distinguish the overlapping fluorescent emissions representative of the two distinct cell populations and longitudinal data reported the discrete pattern of antigen-driven proliferation. Retrieved values were validated both in vitro and in vivo with flow cytometry and significant correlation between all methodologies was achieved. Noninvasive M-SNELS successfully quantified two colocalized fluorescent populations and provides a valid alternative imaging approach to traditional invasive methods for detecting T cell dynamics.

Paper Details

Date Published: 9 July 2012
PDF: 10 pages
J. Biomed. Opt. 17(7) 076013 doi: 10.1117/1.JBO.17.7.076013
Published in: Journal of Biomedical Optics Volume 17, Issue 7
Show Author Affiliations
Rosy Favicchio, Foundation for Research and Technology-Hellas (Greece)
Katerina Oikonomaki, Foundation for Research and Technology-Hellas (Greece)
Kleo Mamalaki, Foundation for Research and Technology-Hellas (Greece)
Giannis Zacharakis, Foundation for Research and Technology-Hellas (Greece)
Athanasios Zacharopoulos, Foundation for Research and Technology-Hellas (Greece)
Jorge Ripoll, Foundation for Research and Technology-Hellas (Greece)

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