Abstract

We report a three-dimensional time-resolved tomographic imaging technique for localizing protein–protein interaction and protein conformational changes in turbid media based on Förster resonant energy-transfer read out using fluorescence lifetime. This application of “tomoFRET” employs an inverse scattering algorithm utilizing the diffusion approximation to the radiative-transfer equation applied to a large tomographic data set of time-gated images. The approach is demonstrated by imaging a highly scattering cylindrical phantom within which are two thin wells containing cytosol preparations of HEK293 cells expressing TN-L15, a cytosolic genetically encoded calcium Förster resonant energy-transfer sensor. A 10mM calcium chloride solution was added to one of the wells, inducing a protein conformation change upon binding to TN-L15, resulting in Förster resonant energy transfer and a corresponding decrease in the donor fluorescence lifetime. We successfully reconstruct spatially resolved maps of the resulting fluorescence lifetime distribution as well as of the quantum efficiency, absorption, and scattering coefficients.

© 2009 Optical Society of America

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  1. J. R. Lakowicz, Principles of Fluorescence Spectroscopy (Plenum, 1999).
  2. D. L. Andrews and D. S. Bradshaw, Eur. J. Phys. 25, 845 (2004).
    [CrossRef]
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    [CrossRef] [PubMed]
  5. V. Y. Soloviev, K. B. Tahir, J. McGinty, D. S. Elson, M. A. A. Neil, P. M. W. French, and S. R. Arridge, Appl. Opt. 46, 7384 (2007).
    [CrossRef] [PubMed]
  6. C. Dunsby, P. M. P. Lanigan, J. McGinty, D. S. Elson, J. Requejo-Isidro, I. Munro, N. Galletly, F. McCann, B. Treanor, B. Onfelt, D. M. Davis, M. A. A. Neil, and P. M. W. French, J. Phys. D 37, 3296 (2004).
    [CrossRef]
  7. J. McGinty, J. Requejo-Isidro, I. Munro, C. B. Talbot, P. A. Kellett, J. D. Hares, C. Dunsby, M. A. A. Neil, and P. M. W. French, J. Phys. D 42, 135103 (2009).
    [CrossRef]
  8. V. Y. Soloviev, C. D'Andrea, G. Valentini, R. Cubeddu, and S. R. Arridge, Appl. Opt. 48, 28 (2009).
    [CrossRef]

2009 (3)

V. Gaind, K. J. Webb, S. Kularatne, and C. A. Bouman, J. Opt. Soc. Am. A 26, 1805 (2009).
[CrossRef]

J. McGinty, J. Requejo-Isidro, I. Munro, C. B. Talbot, P. A. Kellett, J. D. Hares, C. Dunsby, M. A. A. Neil, and P. M. W. French, J. Phys. D 42, 135103 (2009).
[CrossRef]

V. Y. Soloviev, C. D'Andrea, G. Valentini, R. Cubeddu, and S. R. Arridge, Appl. Opt. 48, 28 (2009).
[CrossRef]

2007 (1)

2004 (3)

C. Dunsby, P. M. P. Lanigan, J. McGinty, D. S. Elson, J. Requejo-Isidro, I. Munro, N. Galletly, F. McCann, B. Treanor, B. Onfelt, D. M. Davis, M. A. A. Neil, and P. M. W. French, J. Phys. D 37, 3296 (2004).
[CrossRef]

N. Heim and O. Griesbeck, J. Biol. Chem. 279, 14280 (2004).
[CrossRef] [PubMed]

D. L. Andrews and D. S. Bradshaw, Eur. J. Phys. 25, 845 (2004).
[CrossRef]

1999 (1)

J. R. Lakowicz, Principles of Fluorescence Spectroscopy (Plenum, 1999).

Andrews, D. L.

D. L. Andrews and D. S. Bradshaw, Eur. J. Phys. 25, 845 (2004).
[CrossRef]

Arridge, S. R.

Bouman, C. A.

Bradshaw, D. S.

D. L. Andrews and D. S. Bradshaw, Eur. J. Phys. 25, 845 (2004).
[CrossRef]

Cubeddu, R.

D'Andrea, C.

Davis, D. M.

C. Dunsby, P. M. P. Lanigan, J. McGinty, D. S. Elson, J. Requejo-Isidro, I. Munro, N. Galletly, F. McCann, B. Treanor, B. Onfelt, D. M. Davis, M. A. A. Neil, and P. M. W. French, J. Phys. D 37, 3296 (2004).
[CrossRef]

Dunsby, C.

J. McGinty, J. Requejo-Isidro, I. Munro, C. B. Talbot, P. A. Kellett, J. D. Hares, C. Dunsby, M. A. A. Neil, and P. M. W. French, J. Phys. D 42, 135103 (2009).
[CrossRef]

C. Dunsby, P. M. P. Lanigan, J. McGinty, D. S. Elson, J. Requejo-Isidro, I. Munro, N. Galletly, F. McCann, B. Treanor, B. Onfelt, D. M. Davis, M. A. A. Neil, and P. M. W. French, J. Phys. D 37, 3296 (2004).
[CrossRef]

Elson, D. S.

V. Y. Soloviev, K. B. Tahir, J. McGinty, D. S. Elson, M. A. A. Neil, P. M. W. French, and S. R. Arridge, Appl. Opt. 46, 7384 (2007).
[CrossRef] [PubMed]

C. Dunsby, P. M. P. Lanigan, J. McGinty, D. S. Elson, J. Requejo-Isidro, I. Munro, N. Galletly, F. McCann, B. Treanor, B. Onfelt, D. M. Davis, M. A. A. Neil, and P. M. W. French, J. Phys. D 37, 3296 (2004).
[CrossRef]

French, P. M. W.

J. McGinty, J. Requejo-Isidro, I. Munro, C. B. Talbot, P. A. Kellett, J. D. Hares, C. Dunsby, M. A. A. Neil, and P. M. W. French, J. Phys. D 42, 135103 (2009).
[CrossRef]

V. Y. Soloviev, K. B. Tahir, J. McGinty, D. S. Elson, M. A. A. Neil, P. M. W. French, and S. R. Arridge, Appl. Opt. 46, 7384 (2007).
[CrossRef] [PubMed]

C. Dunsby, P. M. P. Lanigan, J. McGinty, D. S. Elson, J. Requejo-Isidro, I. Munro, N. Galletly, F. McCann, B. Treanor, B. Onfelt, D. M. Davis, M. A. A. Neil, and P. M. W. French, J. Phys. D 37, 3296 (2004).
[CrossRef]

Gaind, V.

Galletly, N.

C. Dunsby, P. M. P. Lanigan, J. McGinty, D. S. Elson, J. Requejo-Isidro, I. Munro, N. Galletly, F. McCann, B. Treanor, B. Onfelt, D. M. Davis, M. A. A. Neil, and P. M. W. French, J. Phys. D 37, 3296 (2004).
[CrossRef]

Griesbeck, O.

N. Heim and O. Griesbeck, J. Biol. Chem. 279, 14280 (2004).
[CrossRef] [PubMed]

Hares, J. D.

J. McGinty, J. Requejo-Isidro, I. Munro, C. B. Talbot, P. A. Kellett, J. D. Hares, C. Dunsby, M. A. A. Neil, and P. M. W. French, J. Phys. D 42, 135103 (2009).
[CrossRef]

Heim, N.

N. Heim and O. Griesbeck, J. Biol. Chem. 279, 14280 (2004).
[CrossRef] [PubMed]

Kellett, P. A.

J. McGinty, J. Requejo-Isidro, I. Munro, C. B. Talbot, P. A. Kellett, J. D. Hares, C. Dunsby, M. A. A. Neil, and P. M. W. French, J. Phys. D 42, 135103 (2009).
[CrossRef]

Kularatne, S.

Lakowicz, J. R.

J. R. Lakowicz, Principles of Fluorescence Spectroscopy (Plenum, 1999).

Lanigan, P. M. P.

C. Dunsby, P. M. P. Lanigan, J. McGinty, D. S. Elson, J. Requejo-Isidro, I. Munro, N. Galletly, F. McCann, B. Treanor, B. Onfelt, D. M. Davis, M. A. A. Neil, and P. M. W. French, J. Phys. D 37, 3296 (2004).
[CrossRef]

McCann, F.

C. Dunsby, P. M. P. Lanigan, J. McGinty, D. S. Elson, J. Requejo-Isidro, I. Munro, N. Galletly, F. McCann, B. Treanor, B. Onfelt, D. M. Davis, M. A. A. Neil, and P. M. W. French, J. Phys. D 37, 3296 (2004).
[CrossRef]

McGinty, J.

J. McGinty, J. Requejo-Isidro, I. Munro, C. B. Talbot, P. A. Kellett, J. D. Hares, C. Dunsby, M. A. A. Neil, and P. M. W. French, J. Phys. D 42, 135103 (2009).
[CrossRef]

V. Y. Soloviev, K. B. Tahir, J. McGinty, D. S. Elson, M. A. A. Neil, P. M. W. French, and S. R. Arridge, Appl. Opt. 46, 7384 (2007).
[CrossRef] [PubMed]

C. Dunsby, P. M. P. Lanigan, J. McGinty, D. S. Elson, J. Requejo-Isidro, I. Munro, N. Galletly, F. McCann, B. Treanor, B. Onfelt, D. M. Davis, M. A. A. Neil, and P. M. W. French, J. Phys. D 37, 3296 (2004).
[CrossRef]

Munro, I.

J. McGinty, J. Requejo-Isidro, I. Munro, C. B. Talbot, P. A. Kellett, J. D. Hares, C. Dunsby, M. A. A. Neil, and P. M. W. French, J. Phys. D 42, 135103 (2009).
[CrossRef]

C. Dunsby, P. M. P. Lanigan, J. McGinty, D. S. Elson, J. Requejo-Isidro, I. Munro, N. Galletly, F. McCann, B. Treanor, B. Onfelt, D. M. Davis, M. A. A. Neil, and P. M. W. French, J. Phys. D 37, 3296 (2004).
[CrossRef]

Neil, M. A. A.

J. McGinty, J. Requejo-Isidro, I. Munro, C. B. Talbot, P. A. Kellett, J. D. Hares, C. Dunsby, M. A. A. Neil, and P. M. W. French, J. Phys. D 42, 135103 (2009).
[CrossRef]

V. Y. Soloviev, K. B. Tahir, J. McGinty, D. S. Elson, M. A. A. Neil, P. M. W. French, and S. R. Arridge, Appl. Opt. 46, 7384 (2007).
[CrossRef] [PubMed]

C. Dunsby, P. M. P. Lanigan, J. McGinty, D. S. Elson, J. Requejo-Isidro, I. Munro, N. Galletly, F. McCann, B. Treanor, B. Onfelt, D. M. Davis, M. A. A. Neil, and P. M. W. French, J. Phys. D 37, 3296 (2004).
[CrossRef]

Onfelt, B.

C. Dunsby, P. M. P. Lanigan, J. McGinty, D. S. Elson, J. Requejo-Isidro, I. Munro, N. Galletly, F. McCann, B. Treanor, B. Onfelt, D. M. Davis, M. A. A. Neil, and P. M. W. French, J. Phys. D 37, 3296 (2004).
[CrossRef]

Requejo-Isidro, J.

J. McGinty, J. Requejo-Isidro, I. Munro, C. B. Talbot, P. A. Kellett, J. D. Hares, C. Dunsby, M. A. A. Neil, and P. M. W. French, J. Phys. D 42, 135103 (2009).
[CrossRef]

C. Dunsby, P. M. P. Lanigan, J. McGinty, D. S. Elson, J. Requejo-Isidro, I. Munro, N. Galletly, F. McCann, B. Treanor, B. Onfelt, D. M. Davis, M. A. A. Neil, and P. M. W. French, J. Phys. D 37, 3296 (2004).
[CrossRef]

Soloviev, V. Y.

Tahir, K. B.

Talbot, C. B.

J. McGinty, J. Requejo-Isidro, I. Munro, C. B. Talbot, P. A. Kellett, J. D. Hares, C. Dunsby, M. A. A. Neil, and P. M. W. French, J. Phys. D 42, 135103 (2009).
[CrossRef]

Treanor, B.

C. Dunsby, P. M. P. Lanigan, J. McGinty, D. S. Elson, J. Requejo-Isidro, I. Munro, N. Galletly, F. McCann, B. Treanor, B. Onfelt, D. M. Davis, M. A. A. Neil, and P. M. W. French, J. Phys. D 37, 3296 (2004).
[CrossRef]

Valentini, G.

Webb, K. J.

Appl. Opt. (2)

Eur. J. Phys. (1)

D. L. Andrews and D. S. Bradshaw, Eur. J. Phys. 25, 845 (2004).
[CrossRef]

J. Biol. Chem. (1)

N. Heim and O. Griesbeck, J. Biol. Chem. 279, 14280 (2004).
[CrossRef] [PubMed]

J. Opt. Soc. Am. A (1)

J. Phys. D (2)

C. Dunsby, P. M. P. Lanigan, J. McGinty, D. S. Elson, J. Requejo-Isidro, I. Munro, N. Galletly, F. McCann, B. Treanor, B. Onfelt, D. M. Davis, M. A. A. Neil, and P. M. W. French, J. Phys. D 37, 3296 (2004).
[CrossRef]

J. McGinty, J. Requejo-Isidro, I. Munro, C. B. Talbot, P. A. Kellett, J. D. Hares, C. Dunsby, M. A. A. Neil, and P. M. W. French, J. Phys. D 42, 135103 (2009).
[CrossRef]

Other (1)

J. R. Lakowicz, Principles of Fluorescence Spectroscopy (Plenum, 1999).

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Figures (2)

Fig. 1
Fig. 1

Fluorescent lifetime image of two wells filled with cytosol preparation containing the FRET sensor placed in a transparent phantom and excited at 436 ± 10 nm . Calcium chloride was added to the right-hand well with a consequent reduction in the CFP fluorescent lifetime.

Fig. 2
Fig. 2

(a) Solid silicone phantom used in experiment. Static mesh adaptation was employed in this study. Slice (b) shows mesh structure. Slices (c)–(f) show reconstruction of fluorescent and optical parameters at phantom’s height y = 40 mm . The quantum yield is shown in (c); distribution of reconstructed lifetime in (e); (d) absorption and (f) diffusion coefficients.

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