Three-dimensional in vivo imaging of green fluorescent protein-expressing T cells in mice with noncontact fluorescence molecular tomography.
Given that optical tomography is capable of quantitatively imaging the distribution of several important chromophores and fluorophores in vivo, there has been a great deal of interest in developing optical imaging systems with increased numbers of measurements under optimal experimental conditions. In this article, we present a novel system that enables three-dimensional imaging of fluorescent probes in whole animals using a noncontact setup, in parallel with a three-dimensional surface reconstruction algorithm. This approach is directed toward the in vivo imaging of fluorophore or fluorescent protein concentration in small animals. The system consists of a rotating sample holder and a lens-coupled charge-coupled device camera in combination with a fiber-coupled laser scanning device. By measuring multiple projections, large data sets can be obtained, thus improving the accuracy of the inversion models used for quantitative three-dimensional reconstruction of fluorochrome distribution, as well as facilitating a higher spatial resolution. In this study, the system was applied to determining the distribution of green fluorescent protein (GFP)-expressing T lymphocytes in a transgenic mouse model, thus demonstrating the potential of the system for studying immune system function. The technique was used to image and reconstruct fluorescence originating from 32 x 10(6) T cells in the thymus and 3 x 10(5) T cells in the spleen.
Garofalakis
Anikitos
A
Institute of Electronic Structure and Laser, Foundation for Research and Techinology Hellas, Heraklion Crete, Greece. agarof@ieslforth.gr
Zacharakis
Giannis
G
Meyer
Heiko
H
Economou
Eleftherios N
EN
Mamalaki
Clio
C
Papamatheakis
Joseph
J
Kioussis
Dimitris
D
Ntziachristos
Vasilis
V
Ripoll
Jorge
J
D000818:Animals; D005434:Flow Cytometry; D005456:Fluorescent Dyes / Q000032:analysis* / Q000378:metabolism; D049452:Green Fluorescent Proteins / Q000032:analysis* / Q000378:metabolism; D021621:Imaging, Three-Dimensional / Q000295:instrumentation / Q000379:methods*; D051379:Mice; D008822:Mice, Transgenic; D013601:T-Lymphocytes / Q000737:chemistry* / Q000276:immunology; D014054:Tomography / Q000295:instrumentation / Q000379:methods*; D051598:Whole Body Imaging / Q000295:instrumentation / Q000379:methods*
D005456:Fluorescent Dyes; D049452:Green Fluorescent Proteins