Research Papers: Imaging

Kinetics of T-cell receptor-dependent antigen recognition determined in vivo by multi-spectral normalized epifluorescence laser scanning

[+] Author Affiliations
Rosy Favicchio

Institute of Molecular Biology and Biotechnology, Foundation for Research and Technology-Hellas, 71110 Heraklion, Crete, Greece

Institute of Electronic Structure and Laser, Foundation for Research and Technology-Hellas, 71110 Heraklion, Crete, Greece

Giannis Zacharakis, Athanasios Zacharopoulos, Jorge Ripoll

Institute of Electronic Structure and Laser, Foundation for Research and Technology-Hellas, 71110 Heraklion, Crete, Greece

Katerina Oikonomaki, Clio Mamalaki

Institute of Molecular Biology and Biotechnology, Foundation for Research and Technology-Hellas, 71110 Heraklion, Crete, Greece

J. Biomed. Opt. 17(7), 076013 (Jul 09, 2012). doi:10.1117/1.JBO.17.7.076013
History: Received December 19, 2011; Revised May 7, 2012; Accepted May 23, 2012
Text Size: A A A

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.

Figures in this Article
© 2012 Society of Photo-Optical Instrumentation Engineers

Citation

Rosy Favicchio ; Giannis Zacharakis ; Katerina Oikonomaki ; Athanasios Zacharopoulos ; Clio Mamalaki, et al.
"Kinetics of T-cell receptor-dependent antigen recognition determined in vivo by multi-spectral normalized epifluorescence laser scanning", J. Biomed. Opt. 17(7), 076013 (Jul 09, 2012). ; http://dx.doi.org/10.1117/1.JBO.17.7.076013


Access This Article
Sign in or Create a personal account to Buy this article ($20 for members, $25 for non-members).

Some tools below are only available to our subscribers or users with an online account.

Related Content

Customize your page view by dragging & repositioning the boxes below.

Related Book Chapters

Topic Collections

Advertisement
  • Don't have an account?
  • Subscribe to the SPIE Digital Library
  • Create a FREE account to sign up for Digital Library content alerts and gain access to institutional subscriptions remotely.
Access This Article
Sign in or Create a personal account to Buy this article ($20 for members, $25 for non-members).
Access This Proceeding
Sign in or Create a personal account to Buy this article ($15 for members, $18 for non-members).
Access This Chapter

Access to SPIE eBooks is limited to subscribing institutions and is not available as part of a personal subscription. Print or electronic versions of individual SPIE books may be purchased via SPIE.org.