Fluorescence lifetime tomography of live cells expressing enhanced green fluorescent protein embedded in a scattering medium exhibiting background autofluorescence.
We present a novel fluorescence lifetime tomography system applied to a highly scattering autofluorescent phantom containing live cells expressing the fluorophore enhanced green fluorescent protein (EGFP). The fluorescence signal was excited using a fiber-laser-pumped supercontinuum source and detected using wide-field time gating imaging. To facilitate rapid 3D reconstruction of the fluorescence lifetime distribution, the time-resolved data were Fourier-transformed in time to give complex functions that formed a data set for the Fourier domain reconstruction. Initially the presence of an unspecified background autofluorescence signal impeded reconstruction of the lifetime distribution, but we show that this problem can be addressed using a simple iterative technique.
Soloviev
Vadim Y
VY
Department of Computer Science, University College London, London WC1E 6BT, UK. v.soloviev@cs.ucl.ac.uk
McGinty
James
J
Tahir
Khadija B
KB
Neil
Mark A
MA
Sardini
Alessandro
A
Hajnal
Joseph V
JV
Arridge
Simon R
SR
French
P M
PM
D000818:Animals; D005453:Fluorescence; D005583:Fourier Analysis; D049452:Green Fluorescent Proteins / Q000737:chemistry*; D007091:Image Processing, Computer-Assisted; D021621:Imaging, Three-Dimensional; D051379:Mice; D015233:Models, Statistical; D041681:NIH 3T3 Cells; D055095:Optics and Photonics; D019047:Phantoms, Imaging; D015203:Reproducibility of Results; D013997:Time Factors; D014054:Tomography / Q000379:methods*
D049452:Green Fluorescent Proteins