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.

[1]  Brian W. Pogue,et al.  Mathematical model for time-resolved and frequency-domain fluorescence spectroscopy in biological tissues. , 1994, Applied optics.

[2]  R. Tsien,et al.  green fluorescent protein , 2020, Catalysis from A to Z.

[3]  Vadim Y Soloviev.  Mesh adaptation technique for Fourier-domain fluorescence lifetime imaging. , 2006, Medical physics.

[4]  Feng Gao,et al.  A linear, featured-data scheme for image reconstruction in time-domain fluorescence molecular tomography. , 2006, Optics express.

[5]  C. Bouman,et al.  Fluorescence optical diffusion tomography. , 2003, Applied optics.

[6]  Vasilis Ntziachristos,et al.  Effects of background fluorescence in fluorescence molecular tomography. , 2005, Applied optics.

[7]  C. L. Hutchinson,et al.  Fluorescence and Absorption Contrast Mechanisms for Biomedical Optical Imaging Using Frequency‐Domain Techniques , 1997, Photochemistry and photobiology.

[8]  Jan Siegel,et al.  Time-domain fluorescence lifetime imaging applied to biological tissue , 2004, Photochemical & photobiological sciences : Official journal of the European Photochemistry Association and the European Society for Photobiology.

[9]  Vasilis Ntziachristos,et al.  Fluorescence molecular tomography in the presence of background fluorescence , 2006, Physics in medicine and biology.

[10]  C A Thompson,et al.  Frequency domain modeling of reradiation in highly scattering media. , 1997, Applied optics.

[11]  D. Boas,et al.  Fluorescence lifetime imaging in turbid media. , 1996, Optics letters.