Abstract

Fluorescence lifetime imaging microscopy (FLIM) has gained popularity as a sensitive technique to monitor the functional/conformational states of reduced nicotinamide adenine dinucleotide (NADH), one of the main compounds of oxidative phosphorylation. In this letter, we apply the technique to characterize the metabolic changes in mouse embryonic fibroblast 3T3 cells upon bacterial infection. A gradual shortening of the decaying time constants in both the short and the long lifetime components of NADH's autofluorescence is detected. The ratio of the short and the long lifetime components' relative contributions, however, shows a rapid increase, indicating the rise of cellular metabolic activity over the course of infection.

© 2010 Chinese Optics Letters

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  1. H. Lodish, A. Berk, C. A. Kaiser, M. Krieger, M. P. Scott, A. Bretscher, H. Ploegh, and P. Matsudaira, Molecular Cell Biology (W. H. Freeman and Co., New York, 2003).
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  3. A. Mousnier, A. Whale, S. Schuller, J. Leong, A. Phillips, and G. Frankel, Infect. Immun. 76, 4669 (2008).
  4. K. Smith, D. Humphreys, P. Hume, and V. Koronakis, Cell Host and Microbe 7, 13 (2010).
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  6. J. Enninga, P. Sansonetti, and R. Tournebize, Trends Microbiol. 15, 483 (2007).
  7. F. Frischknecht, O. Renaud, and S. L. Shorte, Curr. Opin. Microbiol. 9, 297 ( 2006).
  8. B. Kierdaszuk, H. Malak, I. Gryczynski, P. Callis, and J. Lakowicz, Biophys. Chem. 62, (1-3) 1 (1996).
  9. A. Visser and A. Hoek, Photochem. Photobiol. 33, 35(1981).
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  11. V. Ghukasyan and F. J. Kao, J. Phys. Chem. C 113, 11532 (2009).
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  14. V. Ghukasyan, C. C. Hsu, C. R. Liu, F. J. Kao, and T. H. Cheng, J. Biomed. Opt. 15, 016008 (2010).

2010

K. Smith, D. Humphreys, P. Hume, and V. Koronakis, Cell Host and Microbe 7, 13 (2010).

V. Ghukasyan, C. C. Hsu, C. R. Liu, F. J. Kao, and T. H. Cheng, J. Biomed. Opt. 15, 016008 (2010).

2009

V. Ghukasyan and F. J. Kao, J. Phys. Chem. C 113, 11532 (2009).

2008

A. Mousnier, A. Whale, S. Schuller, J. Leong, A. Phillips, and G. Frankel, Infect. Immun. 76, 4669 (2008).

H. W. Wang, V. Ghukasyan, C. T. Chen, Y. H. Wei, H. W. Guo, J. S. Yu, and F. J. Kao, J. Biomed. Opt. 13, 054011 (2008).

2007

J. Enninga, P. Sansonetti, and R. Tournebize, Trends Microbiol. 15, 483 (2007).

V. Ghukasyan, Y. Y. Hsu, S. H. Kung, and F. J. Kao, J. Biomed. Opt. 12, 024016 (2007).

2006

F. Frischknecht, O. Renaud, and S. L. Shorte, Curr. Opin. Microbiol. 9, 297 ( 2006).

2001

M. Abul-Milh, Y. Wu, B. Lau, C. Lingwood, and D. Forster, Infect. Immun. 69, 7356 (2001).

1996

B. Kierdaszuk, H. Malak, I. Gryczynski, P. Callis, and J. Lakowicz, Biophys. Chem. 62, (1-3) 1 (1996).

1981

A. Visser and A. Hoek, Photochem. Photobiol. 33, 35(1981).

Abul-Milh, M.

M. Abul-Milh, Y. Wu, B. Lau, C. Lingwood, and D. Forster, Infect. Immun. 69, 7356 (2001).

Callis, P.

B. Kierdaszuk, H. Malak, I. Gryczynski, P. Callis, and J. Lakowicz, Biophys. Chem. 62, (1-3) 1 (1996).

Chen, C. T.

H. W. Wang, V. Ghukasyan, C. T. Chen, Y. H. Wei, H. W. Guo, J. S. Yu, and F. J. Kao, J. Biomed. Opt. 13, 054011 (2008).

Cheng, T. H.

V. Ghukasyan, C. C. Hsu, C. R. Liu, F. J. Kao, and T. H. Cheng, J. Biomed. Opt. 15, 016008 (2010).

Enninga, J.

J. Enninga, P. Sansonetti, and R. Tournebize, Trends Microbiol. 15, 483 (2007).

Forster, D.

M. Abul-Milh, Y. Wu, B. Lau, C. Lingwood, and D. Forster, Infect. Immun. 69, 7356 (2001).

Frankel, G.

A. Mousnier, A. Whale, S. Schuller, J. Leong, A. Phillips, and G. Frankel, Infect. Immun. 76, 4669 (2008).

Frischknecht, F.

F. Frischknecht, O. Renaud, and S. L. Shorte, Curr. Opin. Microbiol. 9, 297 ( 2006).

Ghukasyan, V.

V. Ghukasyan, C. C. Hsu, C. R. Liu, F. J. Kao, and T. H. Cheng, J. Biomed. Opt. 15, 016008 (2010).

V. Ghukasyan and F. J. Kao, J. Phys. Chem. C 113, 11532 (2009).

H. W. Wang, V. Ghukasyan, C. T. Chen, Y. H. Wei, H. W. Guo, J. S. Yu, and F. J. Kao, J. Biomed. Opt. 13, 054011 (2008).

V. Ghukasyan, Y. Y. Hsu, S. H. Kung, and F. J. Kao, J. Biomed. Opt. 12, 024016 (2007).

Gryczynski, I.

B. Kierdaszuk, H. Malak, I. Gryczynski, P. Callis, and J. Lakowicz, Biophys. Chem. 62, (1-3) 1 (1996).

Guo, H. W.

H. W. Wang, V. Ghukasyan, C. T. Chen, Y. H. Wei, H. W. Guo, J. S. Yu, and F. J. Kao, J. Biomed. Opt. 13, 054011 (2008).

Hoek, A.

A. Visser and A. Hoek, Photochem. Photobiol. 33, 35(1981).

Hsu, C. C.

V. Ghukasyan, C. C. Hsu, C. R. Liu, F. J. Kao, and T. H. Cheng, J. Biomed. Opt. 15, 016008 (2010).

Hsu, Y. Y.

V. Ghukasyan, Y. Y. Hsu, S. H. Kung, and F. J. Kao, J. Biomed. Opt. 12, 024016 (2007).

Hume, P.

K. Smith, D. Humphreys, P. Hume, and V. Koronakis, Cell Host and Microbe 7, 13 (2010).

Humphreys, D.

K. Smith, D. Humphreys, P. Hume, and V. Koronakis, Cell Host and Microbe 7, 13 (2010).

Kao, F. J.

V. Ghukasyan, C. C. Hsu, C. R. Liu, F. J. Kao, and T. H. Cheng, J. Biomed. Opt. 15, 016008 (2010).

V. Ghukasyan and F. J. Kao, J. Phys. Chem. C 113, 11532 (2009).

H. W. Wang, V. Ghukasyan, C. T. Chen, Y. H. Wei, H. W. Guo, J. S. Yu, and F. J. Kao, J. Biomed. Opt. 13, 054011 (2008).

V. Ghukasyan, Y. Y. Hsu, S. H. Kung, and F. J. Kao, J. Biomed. Opt. 12, 024016 (2007).

Kierdaszuk, B.

B. Kierdaszuk, H. Malak, I. Gryczynski, P. Callis, and J. Lakowicz, Biophys. Chem. 62, (1-3) 1 (1996).

Koronakis, V.

K. Smith, D. Humphreys, P. Hume, and V. Koronakis, Cell Host and Microbe 7, 13 (2010).

Kung, S. H.

V. Ghukasyan, Y. Y. Hsu, S. H. Kung, and F. J. Kao, J. Biomed. Opt. 12, 024016 (2007).

Lakowicz, J.

B. Kierdaszuk, H. Malak, I. Gryczynski, P. Callis, and J. Lakowicz, Biophys. Chem. 62, (1-3) 1 (1996).

Lau, B.

M. Abul-Milh, Y. Wu, B. Lau, C. Lingwood, and D. Forster, Infect. Immun. 69, 7356 (2001).

Leong, J.

A. Mousnier, A. Whale, S. Schuller, J. Leong, A. Phillips, and G. Frankel, Infect. Immun. 76, 4669 (2008).

Lingwood, C.

M. Abul-Milh, Y. Wu, B. Lau, C. Lingwood, and D. Forster, Infect. Immun. 69, 7356 (2001).

Liu, C. R.

V. Ghukasyan, C. C. Hsu, C. R. Liu, F. J. Kao, and T. H. Cheng, J. Biomed. Opt. 15, 016008 (2010).

Malak, H.

B. Kierdaszuk, H. Malak, I. Gryczynski, P. Callis, and J. Lakowicz, Biophys. Chem. 62, (1-3) 1 (1996).

Mousnier, A.

A. Mousnier, A. Whale, S. Schuller, J. Leong, A. Phillips, and G. Frankel, Infect. Immun. 76, 4669 (2008).

Phillips, A.

A. Mousnier, A. Whale, S. Schuller, J. Leong, A. Phillips, and G. Frankel, Infect. Immun. 76, 4669 (2008).

Renaud, O.

F. Frischknecht, O. Renaud, and S. L. Shorte, Curr. Opin. Microbiol. 9, 297 ( 2006).

Sansonetti, P.

J. Enninga, P. Sansonetti, and R. Tournebize, Trends Microbiol. 15, 483 (2007).

Schuller, S.

A. Mousnier, A. Whale, S. Schuller, J. Leong, A. Phillips, and G. Frankel, Infect. Immun. 76, 4669 (2008).

Shorte, S. L.

F. Frischknecht, O. Renaud, and S. L. Shorte, Curr. Opin. Microbiol. 9, 297 ( 2006).

Smith, K.

K. Smith, D. Humphreys, P. Hume, and V. Koronakis, Cell Host and Microbe 7, 13 (2010).

Tournebize, R.

J. Enninga, P. Sansonetti, and R. Tournebize, Trends Microbiol. 15, 483 (2007).

Visser, A.

A. Visser and A. Hoek, Photochem. Photobiol. 33, 35(1981).

Wang, H. W.

H. W. Wang, V. Ghukasyan, C. T. Chen, Y. H. Wei, H. W. Guo, J. S. Yu, and F. J. Kao, J. Biomed. Opt. 13, 054011 (2008).

Wei, Y. H.

H. W. Wang, V. Ghukasyan, C. T. Chen, Y. H. Wei, H. W. Guo, J. S. Yu, and F. J. Kao, J. Biomed. Opt. 13, 054011 (2008).

Whale, A.

A. Mousnier, A. Whale, S. Schuller, J. Leong, A. Phillips, and G. Frankel, Infect. Immun. 76, 4669 (2008).

Wu, Y.

M. Abul-Milh, Y. Wu, B. Lau, C. Lingwood, and D. Forster, Infect. Immun. 69, 7356 (2001).

Yu, J. S.

H. W. Wang, V. Ghukasyan, C. T. Chen, Y. H. Wei, H. W. Guo, J. S. Yu, and F. J. Kao, J. Biomed. Opt. 13, 054011 (2008).

Biophys. Chem.

B. Kierdaszuk, H. Malak, I. Gryczynski, P. Callis, and J. Lakowicz, Biophys. Chem. 62, (1-3) 1 (1996).

Cell Host and Microbe

K. Smith, D. Humphreys, P. Hume, and V. Koronakis, Cell Host and Microbe 7, 13 (2010).

Curr. Opin. Microbiol.

F. Frischknecht, O. Renaud, and S. L. Shorte, Curr. Opin. Microbiol. 9, 297 ( 2006).

Infect. Immun.

A. Mousnier, A. Whale, S. Schuller, J. Leong, A. Phillips, and G. Frankel, Infect. Immun. 76, 4669 (2008).

M. Abul-Milh, Y. Wu, B. Lau, C. Lingwood, and D. Forster, Infect. Immun. 69, 7356 (2001).

J. Biomed. Opt.

H. W. Wang, V. Ghukasyan, C. T. Chen, Y. H. Wei, H. W. Guo, J. S. Yu, and F. J. Kao, J. Biomed. Opt. 13, 054011 (2008).

V. Ghukasyan, Y. Y. Hsu, S. H. Kung, and F. J. Kao, J. Biomed. Opt. 12, 024016 (2007).

V. Ghukasyan, C. C. Hsu, C. R. Liu, F. J. Kao, and T. H. Cheng, J. Biomed. Opt. 15, 016008 (2010).

J. Phys. Chem. C

V. Ghukasyan and F. J. Kao, J. Phys. Chem. C 113, 11532 (2009).

Photochem. Photobiol.

A. Visser and A. Hoek, Photochem. Photobiol. 33, 35(1981).

Trends Microbiol.

J. Enninga, P. Sansonetti, and R. Tournebize, Trends Microbiol. 15, 483 (2007).

Other

H. Lodish, A. Berk, C. A. Kaiser, M. Krieger, M. P. Scott, A. Bretscher, H. Ploegh, and P. Matsudaira, Molecular Cell Biology (W. H. Freeman and Co., New York, 2003).

B. Alberts, A. Johnson, J. Lewis, M. Raff, K. Roberts, P. Walter, Molecular Biology of the Cell (Garland Science, New York, 2007).

W. Becker, Advanced Time-Correlated Single Photon Counting Techniques (Springer, Berlin, 2005).

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