Document Detail

Kinetics of T-cell receptor-dependent antigen recognition determined in vivo by multi-spectral normalized epifluorescence laser scanning.
MedLine Citation:
PMID:  22894496     Owner:  NLM     Status:  In-Data-Review    
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.
Rosy Favicchio; Giannis Zacharakis; Katerina Oikonomaki; Athanasios Zacharopoulos; Clio Mamalaki; Jorge Ripoll
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Publication Detail:
Type:  Journal Article    
Journal Detail:
Title:  Journal of biomedical optics     Volume:  17     ISSN:  1560-2281     ISO Abbreviation:  J Biomed Opt     Publication Date:  2012 Jul 
Date Detail:
Created Date:  2012-08-16     Completed Date:  -     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  9605853     Medline TA:  J Biomed Opt     Country:  United States    
Other Details:
Languages:  eng     Pagination:  076013     Citation Subset:  IM    
Institute of Molecular Biology and Biotechnology, Foundation for Research and Technology-Hellas, 71110 Heraklion, Crete, Greece.
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