Abstract

We deduce the signal-to-noise ratio for fluorescence lifetime imaging when using frequency-domain methods. We assume mono-exponential decay and quantum-noise-limited performance. The results are compared with Monte Carlo simulations with good agreement. We also compare our results with previous investigations of time-domain methods for fluorescence lifetime imaging. For a given number of detected photons, we find that frequency-domain and time-domain methods are equally good. The correct choice of detection technique and its parameters is important for obtaining good results.

© 2003 Optical Society of America

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  1. C. G. Morgan, A. C. Mitchell, J. G. Murray, “Nanosecond time-resolved fluorescence microscopy: principles and practice,” Trans. R. Microsc. Soc. 1, 463–466 (1990).
  2. E. P. Buurman, R. Sanders, A. Draaijer, H. C. Gerritsen, J. J. F. van Veen, P. M. Houpt, Y. K. Levine, “Fluorescence lifetime imaging using a confocal laser scanning microscope,” Scanning 14, 155–159 (1992).
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    [CrossRef]
  5. R. Müller, C. Zander, M. Sauer, M. Deimel, D.-S. Ko, S. Siebert, J. Arden-Jacob, G. Deltau, N. J. Marx, K. H. Drexhage, J. Wolfrum, “Time-resolved identification of single molecules in solution with a pulsed semiconductor diode laser,” Chem. Phys. Lett. 262, 716–722 (1996).
    [CrossRef]
  6. A. H. Buist, M. Müller, E. J. Gijsbers, G. J. Brakenhoff, T. S. Sosnowski, T. B. Norris, J. Squier, “Double-pulse fluorescence lifetime measurements,” J. Microsc. (Oxford) 186, 212–220 (1997).
    [CrossRef]
  7. M. Sauer, J. Arden-Jacob, K. H. Drexhage, F. Göbel, U. Lieberwirth, K. Mühlegger, R. Müller, J. Wolfrum, C. Zander, “Time-resolved identification of individual mononucleotide molecules in aqueous solution with pulsed semiconductor lasers,” Bioimaging 6, 14–24 (1998).
    [CrossRef]
  8. J. Sytsma, J. M. Vroom, C. J. de Grauw, H. C. Gerritsen, “Time-gated fluorescence lifetime imaging and microvolume spectroscopy using two-photon excitation,” J. Microsc. (Oxford) 191, 39–51 (1998).
    [CrossRef]
  9. C. J. de Grauw, H. C. Gerritsen, “Multiple time-gate module for fluorescence lifetime imaging,” Appl. Spectrosc. 55, 670–678 (2001).
    [CrossRef]
  10. T. Oida, Y. Sako, A. Kusumi, “Fluorescence lifetime imaging microscopy (flimscopy),” Biophys. J. 64, 676–685 (1993).
    [CrossRef] [PubMed]
  11. C. J. R. van der Oord, H. C. Gerritsen, F. F. G. Rommerts, D. A. Shaw, I. H. Munro, Y. K. Levine, “Micro-volume time-resolved fluorescence spectroscopy using a confocal synchrotron radiation microscope,” Appl. Spectrosc. 49, 1469–1473 (1995).
    [CrossRef]
  12. P. T. C. So, T. French, W. M. Yu, K. M. Berland, C. Y. Dong, E. Gratton, “Time-resolved fluorescence microscopy using two-photon excitation,” Bioimaging 3, 49–63 (1995).
    [CrossRef]
  13. A. Draaijer, R. Sanders, H. C. Gerritsen, “Fluorescence lifetime imaging, a new tool in confocal microscopy,” in Handbook of Biological Confocal Microscopy, J. Pawley, ed. (Plenum, New York, 1995), pp. 491–505.
  14. K. König, P. T. C. So, W. W. Mantulin, B. J. Tromberg, E. Gratton, “Two-photon excited lifetime imaging of autofluorescence in cells during UVA and NIR photostress,” J. Microsc. (Oxford) 183, 197–204 (1996).
  15. H. Brismar, B. Ulfhake, “Fluorescence lifetime measurements in confocal microscopy of neurons labeled with multiple fluorophores,” Nat. Biotechnol. 15, 373–377 (1997).
    [CrossRef] [PubMed]
  16. T. French, P. T. C. So, D. J. Weaver, T. Coelho-Sampaio, E. Gratton, “Two-photon fluorescence lifetime imaging microscopy of macrophage-mediated antigen processing,” J. Microsc. (Oxford) 185, 339–353 (1997).
    [CrossRef]
  17. A. Squire, P. I. H. Bastiaens, “Three dimensional image restoration in fluorescence lifetime imaging microscopy,” J. Microsc. (Oxford) 193, 36–49 (1999).
    [CrossRef]
  18. K. Carlsson, A. Liljeborg, R. M. Andersson, H. Brismar, “Confocal pH imaging of microscopic specimens using fluorescence lifetimes and phase fluorometry: influence of parameter choice on system performance,” J. Microsc. (Oxford) 199, 106–114 (2000).
    [CrossRef]
  19. J. R. Lakowicz, H. Szmacinski, “Fluorescence lifetime-based sensing of pH, Ca+2,K+ and glucose,” Sens. Actuators B 11, 133–143 (1993).
    [CrossRef]
  20. G. Krishnamoorthy, A. Srivastava, “Intracellular dynamics seen through time-resolved fluorescence microscopy,” Curr. Sci. 72, 835–845 (1997).
  21. R. D. Spencer, G. Weber, “Measurements of subnanosecond fluorescence lifetimes with a cross-correlation phase fluorometer,” Ann. N.Y. Acad. Sci. 158, 361–376 (1969).
    [CrossRef]
  22. D. V. O’Connor, D. Phillips, Time-Correlated Single Photon Counting (Academic, New York, 1984).
  23. M. Köllner, J. Wolfrum, “How many photons are necessary for fluorescence lifetime measurements?” Chem. Phys. Lett. 200, 199–204 (1992).
    [CrossRef]
  24. R. M. Ballew, J. N. Demas, “An error analysis of the rapid lifetime determination method for the evaluation of single exponential decays,” Anal. Chem. 61, 30–33 (1989).
    [CrossRef]
  25. E. Gaviola, “Ein Fluorometer. Apparat zur Messung von Fluoreszenzabklingungszeiten,” Z. Phys. 42, 853–861 (1927).
    [CrossRef]
  26. K. Carlsson, A. Liljeborg, “Simultaneous confocal lifetime imaging of multiple fluorophores using the intensity-modulated multiple-wavelength scanning (IMS) technique,” J. Microsc. (Oxford) 191, 119–127 (1998).
    [CrossRef]
  27. K. Carlsson, J. Philip, “Theoretical investigation of the signal-to-noise ratio for different fluorescence lifetime imaging techniques,” in Optical Diagnostics of Living Cells V, D. L. Farkas, R. C. Leif, eds., Proc. SPIE4622, 70–78 (2002).
  28. K. Carlsson, A. Liljeborg, “Confocal fluorescence microscopy using spectral and lifetime information to simultaneously record four fluorophores with high channel separation,” J. Microsc. (Oxford) 185, 37–46 (1997).
    [CrossRef]

2001 (1)

2000 (1)

K. Carlsson, A. Liljeborg, R. M. Andersson, H. Brismar, “Confocal pH imaging of microscopic specimens using fluorescence lifetimes and phase fluorometry: influence of parameter choice on system performance,” J. Microsc. (Oxford) 199, 106–114 (2000).
[CrossRef]

1999 (1)

A. Squire, P. I. H. Bastiaens, “Three dimensional image restoration in fluorescence lifetime imaging microscopy,” J. Microsc. (Oxford) 193, 36–49 (1999).
[CrossRef]

1998 (3)

K. Carlsson, A. Liljeborg, “Simultaneous confocal lifetime imaging of multiple fluorophores using the intensity-modulated multiple-wavelength scanning (IMS) technique,” J. Microsc. (Oxford) 191, 119–127 (1998).
[CrossRef]

M. Sauer, J. Arden-Jacob, K. H. Drexhage, F. Göbel, U. Lieberwirth, K. Mühlegger, R. Müller, J. Wolfrum, C. Zander, “Time-resolved identification of individual mononucleotide molecules in aqueous solution with pulsed semiconductor lasers,” Bioimaging 6, 14–24 (1998).
[CrossRef]

J. Sytsma, J. M. Vroom, C. J. de Grauw, H. C. Gerritsen, “Time-gated fluorescence lifetime imaging and microvolume spectroscopy using two-photon excitation,” J. Microsc. (Oxford) 191, 39–51 (1998).
[CrossRef]

1997 (5)

A. H. Buist, M. Müller, E. J. Gijsbers, G. J. Brakenhoff, T. S. Sosnowski, T. B. Norris, J. Squier, “Double-pulse fluorescence lifetime measurements,” J. Microsc. (Oxford) 186, 212–220 (1997).
[CrossRef]

H. Brismar, B. Ulfhake, “Fluorescence lifetime measurements in confocal microscopy of neurons labeled with multiple fluorophores,” Nat. Biotechnol. 15, 373–377 (1997).
[CrossRef] [PubMed]

T. French, P. T. C. So, D. J. Weaver, T. Coelho-Sampaio, E. Gratton, “Two-photon fluorescence lifetime imaging microscopy of macrophage-mediated antigen processing,” J. Microsc. (Oxford) 185, 339–353 (1997).
[CrossRef]

G. Krishnamoorthy, A. Srivastava, “Intracellular dynamics seen through time-resolved fluorescence microscopy,” Curr. Sci. 72, 835–845 (1997).

K. Carlsson, A. Liljeborg, “Confocal fluorescence microscopy using spectral and lifetime information to simultaneously record four fluorophores with high channel separation,” J. Microsc. (Oxford) 185, 37–46 (1997).
[CrossRef]

1996 (2)

K. König, P. T. C. So, W. W. Mantulin, B. J. Tromberg, E. Gratton, “Two-photon excited lifetime imaging of autofluorescence in cells during UVA and NIR photostress,” J. Microsc. (Oxford) 183, 197–204 (1996).

R. Müller, C. Zander, M. Sauer, M. Deimel, D.-S. Ko, S. Siebert, J. Arden-Jacob, G. Deltau, N. J. Marx, K. H. Drexhage, J. Wolfrum, “Time-resolved identification of single molecules in solution with a pulsed semiconductor diode laser,” Chem. Phys. Lett. 262, 716–722 (1996).
[CrossRef]

1995 (2)

1993 (3)

T. Oida, Y. Sako, A. Kusumi, “Fluorescence lifetime imaging microscopy (flimscopy),” Biophys. J. 64, 676–685 (1993).
[CrossRef] [PubMed]

T. W. J. Gadella, T. M. Jovin, R. M. Clegg, “Fluorescence lifetime imaging microscopy (FLIM): spatial resolution of microstructures on the nanosecond timescale,” Biophys. Chem. 48, 221–239 (1993).
[CrossRef]

J. R. Lakowicz, H. Szmacinski, “Fluorescence lifetime-based sensing of pH, Ca+2,K+ and glucose,” Sens. Actuators B 11, 133–143 (1993).
[CrossRef]

1992 (2)

M. Köllner, J. Wolfrum, “How many photons are necessary for fluorescence lifetime measurements?” Chem. Phys. Lett. 200, 199–204 (1992).
[CrossRef]

E. P. Buurman, R. Sanders, A. Draaijer, H. C. Gerritsen, J. J. F. van Veen, P. M. Houpt, Y. K. Levine, “Fluorescence lifetime imaging using a confocal laser scanning microscope,” Scanning 14, 155–159 (1992).
[CrossRef]

1990 (1)

C. G. Morgan, A. C. Mitchell, J. G. Murray, “Nanosecond time-resolved fluorescence microscopy: principles and practice,” Trans. R. Microsc. Soc. 1, 463–466 (1990).

1989 (1)

R. M. Ballew, J. N. Demas, “An error analysis of the rapid lifetime determination method for the evaluation of single exponential decays,” Anal. Chem. 61, 30–33 (1989).
[CrossRef]

1969 (1)

R. D. Spencer, G. Weber, “Measurements of subnanosecond fluorescence lifetimes with a cross-correlation phase fluorometer,” Ann. N.Y. Acad. Sci. 158, 361–376 (1969).
[CrossRef]

1927 (1)

E. Gaviola, “Ein Fluorometer. Apparat zur Messung von Fluoreszenzabklingungszeiten,” Z. Phys. 42, 853–861 (1927).
[CrossRef]

Andersson, R. M.

K. Carlsson, A. Liljeborg, R. M. Andersson, H. Brismar, “Confocal pH imaging of microscopic specimens using fluorescence lifetimes and phase fluorometry: influence of parameter choice on system performance,” J. Microsc. (Oxford) 199, 106–114 (2000).
[CrossRef]

Arden-Jacob, J.

M. Sauer, J. Arden-Jacob, K. H. Drexhage, F. Göbel, U. Lieberwirth, K. Mühlegger, R. Müller, J. Wolfrum, C. Zander, “Time-resolved identification of individual mononucleotide molecules in aqueous solution with pulsed semiconductor lasers,” Bioimaging 6, 14–24 (1998).
[CrossRef]

R. Müller, C. Zander, M. Sauer, M. Deimel, D.-S. Ko, S. Siebert, J. Arden-Jacob, G. Deltau, N. J. Marx, K. H. Drexhage, J. Wolfrum, “Time-resolved identification of single molecules in solution with a pulsed semiconductor diode laser,” Chem. Phys. Lett. 262, 716–722 (1996).
[CrossRef]

Ballew, R. M.

R. M. Ballew, J. N. Demas, “An error analysis of the rapid lifetime determination method for the evaluation of single exponential decays,” Anal. Chem. 61, 30–33 (1989).
[CrossRef]

Bastiaens, P. I. H.

A. Squire, P. I. H. Bastiaens, “Three dimensional image restoration in fluorescence lifetime imaging microscopy,” J. Microsc. (Oxford) 193, 36–49 (1999).
[CrossRef]

Berland, K. M.

P. T. C. So, T. French, W. M. Yu, K. M. Berland, C. Y. Dong, E. Gratton, “Time-resolved fluorescence microscopy using two-photon excitation,” Bioimaging 3, 49–63 (1995).
[CrossRef]

Brakenhoff, G. J.

A. H. Buist, M. Müller, E. J. Gijsbers, G. J. Brakenhoff, T. S. Sosnowski, T. B. Norris, J. Squier, “Double-pulse fluorescence lifetime measurements,” J. Microsc. (Oxford) 186, 212–220 (1997).
[CrossRef]

Brismar, H.

K. Carlsson, A. Liljeborg, R. M. Andersson, H. Brismar, “Confocal pH imaging of microscopic specimens using fluorescence lifetimes and phase fluorometry: influence of parameter choice on system performance,” J. Microsc. (Oxford) 199, 106–114 (2000).
[CrossRef]

H. Brismar, B. Ulfhake, “Fluorescence lifetime measurements in confocal microscopy of neurons labeled with multiple fluorophores,” Nat. Biotechnol. 15, 373–377 (1997).
[CrossRef] [PubMed]

Buist, A. H.

A. H. Buist, M. Müller, E. J. Gijsbers, G. J. Brakenhoff, T. S. Sosnowski, T. B. Norris, J. Squier, “Double-pulse fluorescence lifetime measurements,” J. Microsc. (Oxford) 186, 212–220 (1997).
[CrossRef]

Buurman, E. P.

E. P. Buurman, R. Sanders, A. Draaijer, H. C. Gerritsen, J. J. F. van Veen, P. M. Houpt, Y. K. Levine, “Fluorescence lifetime imaging using a confocal laser scanning microscope,” Scanning 14, 155–159 (1992).
[CrossRef]

Carlsson, K.

K. Carlsson, A. Liljeborg, R. M. Andersson, H. Brismar, “Confocal pH imaging of microscopic specimens using fluorescence lifetimes and phase fluorometry: influence of parameter choice on system performance,” J. Microsc. (Oxford) 199, 106–114 (2000).
[CrossRef]

K. Carlsson, A. Liljeborg, “Simultaneous confocal lifetime imaging of multiple fluorophores using the intensity-modulated multiple-wavelength scanning (IMS) technique,” J. Microsc. (Oxford) 191, 119–127 (1998).
[CrossRef]

K. Carlsson, A. Liljeborg, “Confocal fluorescence microscopy using spectral and lifetime information to simultaneously record four fluorophores with high channel separation,” J. Microsc. (Oxford) 185, 37–46 (1997).
[CrossRef]

K. Carlsson, J. Philip, “Theoretical investigation of the signal-to-noise ratio for different fluorescence lifetime imaging techniques,” in Optical Diagnostics of Living Cells V, D. L. Farkas, R. C. Leif, eds., Proc. SPIE4622, 70–78 (2002).

Clegg, R. M.

T. W. J. Gadella, T. M. Jovin, R. M. Clegg, “Fluorescence lifetime imaging microscopy (FLIM): spatial resolution of microstructures on the nanosecond timescale,” Biophys. Chem. 48, 221–239 (1993).
[CrossRef]

Coelho-Sampaio, T.

T. French, P. T. C. So, D. J. Weaver, T. Coelho-Sampaio, E. Gratton, “Two-photon fluorescence lifetime imaging microscopy of macrophage-mediated antigen processing,” J. Microsc. (Oxford) 185, 339–353 (1997).
[CrossRef]

de Grauw, C. J.

C. J. de Grauw, H. C. Gerritsen, “Multiple time-gate module for fluorescence lifetime imaging,” Appl. Spectrosc. 55, 670–678 (2001).
[CrossRef]

J. Sytsma, J. M. Vroom, C. J. de Grauw, H. C. Gerritsen, “Time-gated fluorescence lifetime imaging and microvolume spectroscopy using two-photon excitation,” J. Microsc. (Oxford) 191, 39–51 (1998).
[CrossRef]

Deimel, M.

R. Müller, C. Zander, M. Sauer, M. Deimel, D.-S. Ko, S. Siebert, J. Arden-Jacob, G. Deltau, N. J. Marx, K. H. Drexhage, J. Wolfrum, “Time-resolved identification of single molecules in solution with a pulsed semiconductor diode laser,” Chem. Phys. Lett. 262, 716–722 (1996).
[CrossRef]

Deltau, G.

R. Müller, C. Zander, M. Sauer, M. Deimel, D.-S. Ko, S. Siebert, J. Arden-Jacob, G. Deltau, N. J. Marx, K. H. Drexhage, J. Wolfrum, “Time-resolved identification of single molecules in solution with a pulsed semiconductor diode laser,” Chem. Phys. Lett. 262, 716–722 (1996).
[CrossRef]

Demas, J. N.

R. M. Ballew, J. N. Demas, “An error analysis of the rapid lifetime determination method for the evaluation of single exponential decays,” Anal. Chem. 61, 30–33 (1989).
[CrossRef]

Dong, C. Y.

P. T. C. So, T. French, W. M. Yu, K. M. Berland, C. Y. Dong, E. Gratton, “Time-resolved fluorescence microscopy using two-photon excitation,” Bioimaging 3, 49–63 (1995).
[CrossRef]

Draaijer, A.

E. P. Buurman, R. Sanders, A. Draaijer, H. C. Gerritsen, J. J. F. van Veen, P. M. Houpt, Y. K. Levine, “Fluorescence lifetime imaging using a confocal laser scanning microscope,” Scanning 14, 155–159 (1992).
[CrossRef]

A. Draaijer, R. Sanders, H. C. Gerritsen, “Fluorescence lifetime imaging, a new tool in confocal microscopy,” in Handbook of Biological Confocal Microscopy, J. Pawley, ed. (Plenum, New York, 1995), pp. 491–505.

Drexhage, K. H.

M. Sauer, J. Arden-Jacob, K. H. Drexhage, F. Göbel, U. Lieberwirth, K. Mühlegger, R. Müller, J. Wolfrum, C. Zander, “Time-resolved identification of individual mononucleotide molecules in aqueous solution with pulsed semiconductor lasers,” Bioimaging 6, 14–24 (1998).
[CrossRef]

R. Müller, C. Zander, M. Sauer, M. Deimel, D.-S. Ko, S. Siebert, J. Arden-Jacob, G. Deltau, N. J. Marx, K. H. Drexhage, J. Wolfrum, “Time-resolved identification of single molecules in solution with a pulsed semiconductor diode laser,” Chem. Phys. Lett. 262, 716–722 (1996).
[CrossRef]

French, T.

T. French, P. T. C. So, D. J. Weaver, T. Coelho-Sampaio, E. Gratton, “Two-photon fluorescence lifetime imaging microscopy of macrophage-mediated antigen processing,” J. Microsc. (Oxford) 185, 339–353 (1997).
[CrossRef]

P. T. C. So, T. French, W. M. Yu, K. M. Berland, C. Y. Dong, E. Gratton, “Time-resolved fluorescence microscopy using two-photon excitation,” Bioimaging 3, 49–63 (1995).
[CrossRef]

Gadella, T. W. J.

T. W. J. Gadella, T. M. Jovin, R. M. Clegg, “Fluorescence lifetime imaging microscopy (FLIM): spatial resolution of microstructures on the nanosecond timescale,” Biophys. Chem. 48, 221–239 (1993).
[CrossRef]

Gaviola, E.

E. Gaviola, “Ein Fluorometer. Apparat zur Messung von Fluoreszenzabklingungszeiten,” Z. Phys. 42, 853–861 (1927).
[CrossRef]

Gerritsen, H. C.

C. J. de Grauw, H. C. Gerritsen, “Multiple time-gate module for fluorescence lifetime imaging,” Appl. Spectrosc. 55, 670–678 (2001).
[CrossRef]

J. Sytsma, J. M. Vroom, C. J. de Grauw, H. C. Gerritsen, “Time-gated fluorescence lifetime imaging and microvolume spectroscopy using two-photon excitation,” J. Microsc. (Oxford) 191, 39–51 (1998).
[CrossRef]

C. J. R. van der Oord, H. C. Gerritsen, F. F. G. Rommerts, D. A. Shaw, I. H. Munro, Y. K. Levine, “Micro-volume time-resolved fluorescence spectroscopy using a confocal synchrotron radiation microscope,” Appl. Spectrosc. 49, 1469–1473 (1995).
[CrossRef]

E. P. Buurman, R. Sanders, A. Draaijer, H. C. Gerritsen, J. J. F. van Veen, P. M. Houpt, Y. K. Levine, “Fluorescence lifetime imaging using a confocal laser scanning microscope,” Scanning 14, 155–159 (1992).
[CrossRef]

A. Draaijer, R. Sanders, H. C. Gerritsen, “Fluorescence lifetime imaging, a new tool in confocal microscopy,” in Handbook of Biological Confocal Microscopy, J. Pawley, ed. (Plenum, New York, 1995), pp. 491–505.

Gijsbers, E. J.

A. H. Buist, M. Müller, E. J. Gijsbers, G. J. Brakenhoff, T. S. Sosnowski, T. B. Norris, J. Squier, “Double-pulse fluorescence lifetime measurements,” J. Microsc. (Oxford) 186, 212–220 (1997).
[CrossRef]

Göbel, F.

M. Sauer, J. Arden-Jacob, K. H. Drexhage, F. Göbel, U. Lieberwirth, K. Mühlegger, R. Müller, J. Wolfrum, C. Zander, “Time-resolved identification of individual mononucleotide molecules in aqueous solution with pulsed semiconductor lasers,” Bioimaging 6, 14–24 (1998).
[CrossRef]

Gratton, E.

T. French, P. T. C. So, D. J. Weaver, T. Coelho-Sampaio, E. Gratton, “Two-photon fluorescence lifetime imaging microscopy of macrophage-mediated antigen processing,” J. Microsc. (Oxford) 185, 339–353 (1997).
[CrossRef]

K. König, P. T. C. So, W. W. Mantulin, B. J. Tromberg, E. Gratton, “Two-photon excited lifetime imaging of autofluorescence in cells during UVA and NIR photostress,” J. Microsc. (Oxford) 183, 197–204 (1996).

P. T. C. So, T. French, W. M. Yu, K. M. Berland, C. Y. Dong, E. Gratton, “Time-resolved fluorescence microscopy using two-photon excitation,” Bioimaging 3, 49–63 (1995).
[CrossRef]

Houpt, P. M.

E. P. Buurman, R. Sanders, A. Draaijer, H. C. Gerritsen, J. J. F. van Veen, P. M. Houpt, Y. K. Levine, “Fluorescence lifetime imaging using a confocal laser scanning microscope,” Scanning 14, 155–159 (1992).
[CrossRef]

Jovin, T. M.

T. W. J. Gadella, T. M. Jovin, R. M. Clegg, “Fluorescence lifetime imaging microscopy (FLIM): spatial resolution of microstructures on the nanosecond timescale,” Biophys. Chem. 48, 221–239 (1993).
[CrossRef]

Ko, D.-S.

R. Müller, C. Zander, M. Sauer, M. Deimel, D.-S. Ko, S. Siebert, J. Arden-Jacob, G. Deltau, N. J. Marx, K. H. Drexhage, J. Wolfrum, “Time-resolved identification of single molecules in solution with a pulsed semiconductor diode laser,” Chem. Phys. Lett. 262, 716–722 (1996).
[CrossRef]

Köllner, M.

M. Köllner, J. Wolfrum, “How many photons are necessary for fluorescence lifetime measurements?” Chem. Phys. Lett. 200, 199–204 (1992).
[CrossRef]

König, K.

K. König, P. T. C. So, W. W. Mantulin, B. J. Tromberg, E. Gratton, “Two-photon excited lifetime imaging of autofluorescence in cells during UVA and NIR photostress,” J. Microsc. (Oxford) 183, 197–204 (1996).

Krishnamoorthy, G.

G. Krishnamoorthy, A. Srivastava, “Intracellular dynamics seen through time-resolved fluorescence microscopy,” Curr. Sci. 72, 835–845 (1997).

Kusumi, A.

T. Oida, Y. Sako, A. Kusumi, “Fluorescence lifetime imaging microscopy (flimscopy),” Biophys. J. 64, 676–685 (1993).
[CrossRef] [PubMed]

Lakowicz, J. R.

J. R. Lakowicz, H. Szmacinski, “Fluorescence lifetime-based sensing of pH, Ca+2,K+ and glucose,” Sens. Actuators B 11, 133–143 (1993).
[CrossRef]

Levine, Y. K.

C. J. R. van der Oord, H. C. Gerritsen, F. F. G. Rommerts, D. A. Shaw, I. H. Munro, Y. K. Levine, “Micro-volume time-resolved fluorescence spectroscopy using a confocal synchrotron radiation microscope,” Appl. Spectrosc. 49, 1469–1473 (1995).
[CrossRef]

E. P. Buurman, R. Sanders, A. Draaijer, H. C. Gerritsen, J. J. F. van Veen, P. M. Houpt, Y. K. Levine, “Fluorescence lifetime imaging using a confocal laser scanning microscope,” Scanning 14, 155–159 (1992).
[CrossRef]

Lieberwirth, U.

M. Sauer, J. Arden-Jacob, K. H. Drexhage, F. Göbel, U. Lieberwirth, K. Mühlegger, R. Müller, J. Wolfrum, C. Zander, “Time-resolved identification of individual mononucleotide molecules in aqueous solution with pulsed semiconductor lasers,” Bioimaging 6, 14–24 (1998).
[CrossRef]

Liljeborg, A.

K. Carlsson, A. Liljeborg, R. M. Andersson, H. Brismar, “Confocal pH imaging of microscopic specimens using fluorescence lifetimes and phase fluorometry: influence of parameter choice on system performance,” J. Microsc. (Oxford) 199, 106–114 (2000).
[CrossRef]

K. Carlsson, A. Liljeborg, “Simultaneous confocal lifetime imaging of multiple fluorophores using the intensity-modulated multiple-wavelength scanning (IMS) technique,” J. Microsc. (Oxford) 191, 119–127 (1998).
[CrossRef]

K. Carlsson, A. Liljeborg, “Confocal fluorescence microscopy using spectral and lifetime information to simultaneously record four fluorophores with high channel separation,” J. Microsc. (Oxford) 185, 37–46 (1997).
[CrossRef]

Mantulin, W. W.

K. König, P. T. C. So, W. W. Mantulin, B. J. Tromberg, E. Gratton, “Two-photon excited lifetime imaging of autofluorescence in cells during UVA and NIR photostress,” J. Microsc. (Oxford) 183, 197–204 (1996).

Marx, N. J.

R. Müller, C. Zander, M. Sauer, M. Deimel, D.-S. Ko, S. Siebert, J. Arden-Jacob, G. Deltau, N. J. Marx, K. H. Drexhage, J. Wolfrum, “Time-resolved identification of single molecules in solution with a pulsed semiconductor diode laser,” Chem. Phys. Lett. 262, 716–722 (1996).
[CrossRef]

Mitchell, A. C.

C. G. Morgan, A. C. Mitchell, J. G. Murray, “Nanosecond time-resolved fluorescence microscopy: principles and practice,” Trans. R. Microsc. Soc. 1, 463–466 (1990).

Morgan, C. G.

C. G. Morgan, A. C. Mitchell, J. G. Murray, “Nanosecond time-resolved fluorescence microscopy: principles and practice,” Trans. R. Microsc. Soc. 1, 463–466 (1990).

Mühlegger, K.

M. Sauer, J. Arden-Jacob, K. H. Drexhage, F. Göbel, U. Lieberwirth, K. Mühlegger, R. Müller, J. Wolfrum, C. Zander, “Time-resolved identification of individual mononucleotide molecules in aqueous solution with pulsed semiconductor lasers,” Bioimaging 6, 14–24 (1998).
[CrossRef]

Müller, M.

A. H. Buist, M. Müller, E. J. Gijsbers, G. J. Brakenhoff, T. S. Sosnowski, T. B. Norris, J. Squier, “Double-pulse fluorescence lifetime measurements,” J. Microsc. (Oxford) 186, 212–220 (1997).
[CrossRef]

Müller, R.

M. Sauer, J. Arden-Jacob, K. H. Drexhage, F. Göbel, U. Lieberwirth, K. Mühlegger, R. Müller, J. Wolfrum, C. Zander, “Time-resolved identification of individual mononucleotide molecules in aqueous solution with pulsed semiconductor lasers,” Bioimaging 6, 14–24 (1998).
[CrossRef]

R. Müller, C. Zander, M. Sauer, M. Deimel, D.-S. Ko, S. Siebert, J. Arden-Jacob, G. Deltau, N. J. Marx, K. H. Drexhage, J. Wolfrum, “Time-resolved identification of single molecules in solution with a pulsed semiconductor diode laser,” Chem. Phys. Lett. 262, 716–722 (1996).
[CrossRef]

Munro, I. H.

Murray, J. G.

C. G. Morgan, A. C. Mitchell, J. G. Murray, “Nanosecond time-resolved fluorescence microscopy: principles and practice,” Trans. R. Microsc. Soc. 1, 463–466 (1990).

Norris, T. B.

A. H. Buist, M. Müller, E. J. Gijsbers, G. J. Brakenhoff, T. S. Sosnowski, T. B. Norris, J. Squier, “Double-pulse fluorescence lifetime measurements,” J. Microsc. (Oxford) 186, 212–220 (1997).
[CrossRef]

O’Connor, D. V.

D. V. O’Connor, D. Phillips, Time-Correlated Single Photon Counting (Academic, New York, 1984).

Oida, T.

T. Oida, Y. Sako, A. Kusumi, “Fluorescence lifetime imaging microscopy (flimscopy),” Biophys. J. 64, 676–685 (1993).
[CrossRef] [PubMed]

Philip, J.

K. Carlsson, J. Philip, “Theoretical investigation of the signal-to-noise ratio for different fluorescence lifetime imaging techniques,” in Optical Diagnostics of Living Cells V, D. L. Farkas, R. C. Leif, eds., Proc. SPIE4622, 70–78 (2002).

Phillips, D.

D. V. O’Connor, D. Phillips, Time-Correlated Single Photon Counting (Academic, New York, 1984).

Piston, D. W.

D. W. Piston, D. R. Sandison, W. W. Webb, “Time-resolved fluorescence imaging and background rejection by two-photon excitation in laser scanning microscopy,” in Time-Resolved Laser Spectroscopy in Biochemistry III, J. R. Lakowicz, ed., Proc. SPIE1640, 379–389 (1992).

Rommerts, F. F. G.

Sako, Y.

T. Oida, Y. Sako, A. Kusumi, “Fluorescence lifetime imaging microscopy (flimscopy),” Biophys. J. 64, 676–685 (1993).
[CrossRef] [PubMed]

Sanders, R.

E. P. Buurman, R. Sanders, A. Draaijer, H. C. Gerritsen, J. J. F. van Veen, P. M. Houpt, Y. K. Levine, “Fluorescence lifetime imaging using a confocal laser scanning microscope,” Scanning 14, 155–159 (1992).
[CrossRef]

A. Draaijer, R. Sanders, H. C. Gerritsen, “Fluorescence lifetime imaging, a new tool in confocal microscopy,” in Handbook of Biological Confocal Microscopy, J. Pawley, ed. (Plenum, New York, 1995), pp. 491–505.

Sandison, D. R.

D. W. Piston, D. R. Sandison, W. W. Webb, “Time-resolved fluorescence imaging and background rejection by two-photon excitation in laser scanning microscopy,” in Time-Resolved Laser Spectroscopy in Biochemistry III, J. R. Lakowicz, ed., Proc. SPIE1640, 379–389 (1992).

Sauer, M.

M. Sauer, J. Arden-Jacob, K. H. Drexhage, F. Göbel, U. Lieberwirth, K. Mühlegger, R. Müller, J. Wolfrum, C. Zander, “Time-resolved identification of individual mononucleotide molecules in aqueous solution with pulsed semiconductor lasers,” Bioimaging 6, 14–24 (1998).
[CrossRef]

R. Müller, C. Zander, M. Sauer, M. Deimel, D.-S. Ko, S. Siebert, J. Arden-Jacob, G. Deltau, N. J. Marx, K. H. Drexhage, J. Wolfrum, “Time-resolved identification of single molecules in solution with a pulsed semiconductor diode laser,” Chem. Phys. Lett. 262, 716–722 (1996).
[CrossRef]

Shaw, D. A.

Siebert, S.

R. Müller, C. Zander, M. Sauer, M. Deimel, D.-S. Ko, S. Siebert, J. Arden-Jacob, G. Deltau, N. J. Marx, K. H. Drexhage, J. Wolfrum, “Time-resolved identification of single molecules in solution with a pulsed semiconductor diode laser,” Chem. Phys. Lett. 262, 716–722 (1996).
[CrossRef]

So, P. T. C.

T. French, P. T. C. So, D. J. Weaver, T. Coelho-Sampaio, E. Gratton, “Two-photon fluorescence lifetime imaging microscopy of macrophage-mediated antigen processing,” J. Microsc. (Oxford) 185, 339–353 (1997).
[CrossRef]

K. König, P. T. C. So, W. W. Mantulin, B. J. Tromberg, E. Gratton, “Two-photon excited lifetime imaging of autofluorescence in cells during UVA and NIR photostress,” J. Microsc. (Oxford) 183, 197–204 (1996).

P. T. C. So, T. French, W. M. Yu, K. M. Berland, C. Y. Dong, E. Gratton, “Time-resolved fluorescence microscopy using two-photon excitation,” Bioimaging 3, 49–63 (1995).
[CrossRef]

Sosnowski, T. S.

A. H. Buist, M. Müller, E. J. Gijsbers, G. J. Brakenhoff, T. S. Sosnowski, T. B. Norris, J. Squier, “Double-pulse fluorescence lifetime measurements,” J. Microsc. (Oxford) 186, 212–220 (1997).
[CrossRef]

Spencer, R. D.

R. D. Spencer, G. Weber, “Measurements of subnanosecond fluorescence lifetimes with a cross-correlation phase fluorometer,” Ann. N.Y. Acad. Sci. 158, 361–376 (1969).
[CrossRef]

Squier, J.

A. H. Buist, M. Müller, E. J. Gijsbers, G. J. Brakenhoff, T. S. Sosnowski, T. B. Norris, J. Squier, “Double-pulse fluorescence lifetime measurements,” J. Microsc. (Oxford) 186, 212–220 (1997).
[CrossRef]

Squire, A.

A. Squire, P. I. H. Bastiaens, “Three dimensional image restoration in fluorescence lifetime imaging microscopy,” J. Microsc. (Oxford) 193, 36–49 (1999).
[CrossRef]

Srivastava, A.

G. Krishnamoorthy, A. Srivastava, “Intracellular dynamics seen through time-resolved fluorescence microscopy,” Curr. Sci. 72, 835–845 (1997).

Sytsma, J.

J. Sytsma, J. M. Vroom, C. J. de Grauw, H. C. Gerritsen, “Time-gated fluorescence lifetime imaging and microvolume spectroscopy using two-photon excitation,” J. Microsc. (Oxford) 191, 39–51 (1998).
[CrossRef]

Szmacinski, H.

J. R. Lakowicz, H. Szmacinski, “Fluorescence lifetime-based sensing of pH, Ca+2,K+ and glucose,” Sens. Actuators B 11, 133–143 (1993).
[CrossRef]

Tromberg, B. J.

K. König, P. T. C. So, W. W. Mantulin, B. J. Tromberg, E. Gratton, “Two-photon excited lifetime imaging of autofluorescence in cells during UVA and NIR photostress,” J. Microsc. (Oxford) 183, 197–204 (1996).

Ulfhake, B.

H. Brismar, B. Ulfhake, “Fluorescence lifetime measurements in confocal microscopy of neurons labeled with multiple fluorophores,” Nat. Biotechnol. 15, 373–377 (1997).
[CrossRef] [PubMed]

van der Oord, C. J. R.

van Veen, J. J. F.

E. P. Buurman, R. Sanders, A. Draaijer, H. C. Gerritsen, J. J. F. van Veen, P. M. Houpt, Y. K. Levine, “Fluorescence lifetime imaging using a confocal laser scanning microscope,” Scanning 14, 155–159 (1992).
[CrossRef]

Vroom, J. M.

J. Sytsma, J. M. Vroom, C. J. de Grauw, H. C. Gerritsen, “Time-gated fluorescence lifetime imaging and microvolume spectroscopy using two-photon excitation,” J. Microsc. (Oxford) 191, 39–51 (1998).
[CrossRef]

Weaver, D. J.

T. French, P. T. C. So, D. J. Weaver, T. Coelho-Sampaio, E. Gratton, “Two-photon fluorescence lifetime imaging microscopy of macrophage-mediated antigen processing,” J. Microsc. (Oxford) 185, 339–353 (1997).
[CrossRef]

Webb, W. W.

D. W. Piston, D. R. Sandison, W. W. Webb, “Time-resolved fluorescence imaging and background rejection by two-photon excitation in laser scanning microscopy,” in Time-Resolved Laser Spectroscopy in Biochemistry III, J. R. Lakowicz, ed., Proc. SPIE1640, 379–389 (1992).

Weber, G.

R. D. Spencer, G. Weber, “Measurements of subnanosecond fluorescence lifetimes with a cross-correlation phase fluorometer,” Ann. N.Y. Acad. Sci. 158, 361–376 (1969).
[CrossRef]

Wolfrum, J.

M. Sauer, J. Arden-Jacob, K. H. Drexhage, F. Göbel, U. Lieberwirth, K. Mühlegger, R. Müller, J. Wolfrum, C. Zander, “Time-resolved identification of individual mononucleotide molecules in aqueous solution with pulsed semiconductor lasers,” Bioimaging 6, 14–24 (1998).
[CrossRef]

R. Müller, C. Zander, M. Sauer, M. Deimel, D.-S. Ko, S. Siebert, J. Arden-Jacob, G. Deltau, N. J. Marx, K. H. Drexhage, J. Wolfrum, “Time-resolved identification of single molecules in solution with a pulsed semiconductor diode laser,” Chem. Phys. Lett. 262, 716–722 (1996).
[CrossRef]

M. Köllner, J. Wolfrum, “How many photons are necessary for fluorescence lifetime measurements?” Chem. Phys. Lett. 200, 199–204 (1992).
[CrossRef]

Yu, W. M.

P. T. C. So, T. French, W. M. Yu, K. M. Berland, C. Y. Dong, E. Gratton, “Time-resolved fluorescence microscopy using two-photon excitation,” Bioimaging 3, 49–63 (1995).
[CrossRef]

Zander, C.

M. Sauer, J. Arden-Jacob, K. H. Drexhage, F. Göbel, U. Lieberwirth, K. Mühlegger, R. Müller, J. Wolfrum, C. Zander, “Time-resolved identification of individual mononucleotide molecules in aqueous solution with pulsed semiconductor lasers,” Bioimaging 6, 14–24 (1998).
[CrossRef]

R. Müller, C. Zander, M. Sauer, M. Deimel, D.-S. Ko, S. Siebert, J. Arden-Jacob, G. Deltau, N. J. Marx, K. H. Drexhage, J. Wolfrum, “Time-resolved identification of single molecules in solution with a pulsed semiconductor diode laser,” Chem. Phys. Lett. 262, 716–722 (1996).
[CrossRef]

Anal. Chem. (1)

R. M. Ballew, J. N. Demas, “An error analysis of the rapid lifetime determination method for the evaluation of single exponential decays,” Anal. Chem. 61, 30–33 (1989).
[CrossRef]

Ann. N.Y. Acad. Sci. (1)

R. D. Spencer, G. Weber, “Measurements of subnanosecond fluorescence lifetimes with a cross-correlation phase fluorometer,” Ann. N.Y. Acad. Sci. 158, 361–376 (1969).
[CrossRef]

Appl. Spectrosc. (2)

Bioimaging (2)

M. Sauer, J. Arden-Jacob, K. H. Drexhage, F. Göbel, U. Lieberwirth, K. Mühlegger, R. Müller, J. Wolfrum, C. Zander, “Time-resolved identification of individual mononucleotide molecules in aqueous solution with pulsed semiconductor lasers,” Bioimaging 6, 14–24 (1998).
[CrossRef]

P. T. C. So, T. French, W. M. Yu, K. M. Berland, C. Y. Dong, E. Gratton, “Time-resolved fluorescence microscopy using two-photon excitation,” Bioimaging 3, 49–63 (1995).
[CrossRef]

Biophys. Chem. (1)

T. W. J. Gadella, T. M. Jovin, R. M. Clegg, “Fluorescence lifetime imaging microscopy (FLIM): spatial resolution of microstructures on the nanosecond timescale,” Biophys. Chem. 48, 221–239 (1993).
[CrossRef]

Biophys. J. (1)

T. Oida, Y. Sako, A. Kusumi, “Fluorescence lifetime imaging microscopy (flimscopy),” Biophys. J. 64, 676–685 (1993).
[CrossRef] [PubMed]

Chem. Phys. Lett. (2)

R. Müller, C. Zander, M. Sauer, M. Deimel, D.-S. Ko, S. Siebert, J. Arden-Jacob, G. Deltau, N. J. Marx, K. H. Drexhage, J. Wolfrum, “Time-resolved identification of single molecules in solution with a pulsed semiconductor diode laser,” Chem. Phys. Lett. 262, 716–722 (1996).
[CrossRef]

M. Köllner, J. Wolfrum, “How many photons are necessary for fluorescence lifetime measurements?” Chem. Phys. Lett. 200, 199–204 (1992).
[CrossRef]

Curr. Sci. (1)

G. Krishnamoorthy, A. Srivastava, “Intracellular dynamics seen through time-resolved fluorescence microscopy,” Curr. Sci. 72, 835–845 (1997).

J. Microsc. (Oxford) (8)

K. Carlsson, A. Liljeborg, “Simultaneous confocal lifetime imaging of multiple fluorophores using the intensity-modulated multiple-wavelength scanning (IMS) technique,” J. Microsc. (Oxford) 191, 119–127 (1998).
[CrossRef]

K. König, P. T. C. So, W. W. Mantulin, B. J. Tromberg, E. Gratton, “Two-photon excited lifetime imaging of autofluorescence in cells during UVA and NIR photostress,” J. Microsc. (Oxford) 183, 197–204 (1996).

T. French, P. T. C. So, D. J. Weaver, T. Coelho-Sampaio, E. Gratton, “Two-photon fluorescence lifetime imaging microscopy of macrophage-mediated antigen processing,” J. Microsc. (Oxford) 185, 339–353 (1997).
[CrossRef]

A. Squire, P. I. H. Bastiaens, “Three dimensional image restoration in fluorescence lifetime imaging microscopy,” J. Microsc. (Oxford) 193, 36–49 (1999).
[CrossRef]

K. Carlsson, A. Liljeborg, R. M. Andersson, H. Brismar, “Confocal pH imaging of microscopic specimens using fluorescence lifetimes and phase fluorometry: influence of parameter choice on system performance,” J. Microsc. (Oxford) 199, 106–114 (2000).
[CrossRef]

A. H. Buist, M. Müller, E. J. Gijsbers, G. J. Brakenhoff, T. S. Sosnowski, T. B. Norris, J. Squier, “Double-pulse fluorescence lifetime measurements,” J. Microsc. (Oxford) 186, 212–220 (1997).
[CrossRef]

J. Sytsma, J. M. Vroom, C. J. de Grauw, H. C. Gerritsen, “Time-gated fluorescence lifetime imaging and microvolume spectroscopy using two-photon excitation,” J. Microsc. (Oxford) 191, 39–51 (1998).
[CrossRef]

K. Carlsson, A. Liljeborg, “Confocal fluorescence microscopy using spectral and lifetime information to simultaneously record four fluorophores with high channel separation,” J. Microsc. (Oxford) 185, 37–46 (1997).
[CrossRef]

Nat. Biotechnol. (1)

H. Brismar, B. Ulfhake, “Fluorescence lifetime measurements in confocal microscopy of neurons labeled with multiple fluorophores,” Nat. Biotechnol. 15, 373–377 (1997).
[CrossRef] [PubMed]

Scanning (1)

E. P. Buurman, R. Sanders, A. Draaijer, H. C. Gerritsen, J. J. F. van Veen, P. M. Houpt, Y. K. Levine, “Fluorescence lifetime imaging using a confocal laser scanning microscope,” Scanning 14, 155–159 (1992).
[CrossRef]

Sens. Actuators B (1)

J. R. Lakowicz, H. Szmacinski, “Fluorescence lifetime-based sensing of pH, Ca+2,K+ and glucose,” Sens. Actuators B 11, 133–143 (1993).
[CrossRef]

Trans. R. Microsc. Soc. (1)

C. G. Morgan, A. C. Mitchell, J. G. Murray, “Nanosecond time-resolved fluorescence microscopy: principles and practice,” Trans. R. Microsc. Soc. 1, 463–466 (1990).

Z. Phys. (1)

E. Gaviola, “Ein Fluorometer. Apparat zur Messung von Fluoreszenzabklingungszeiten,” Z. Phys. 42, 853–861 (1927).
[CrossRef]

Other (4)

K. Carlsson, J. Philip, “Theoretical investigation of the signal-to-noise ratio for different fluorescence lifetime imaging techniques,” in Optical Diagnostics of Living Cells V, D. L. Farkas, R. C. Leif, eds., Proc. SPIE4622, 70–78 (2002).

D. V. O’Connor, D. Phillips, Time-Correlated Single Photon Counting (Academic, New York, 1984).

A. Draaijer, R. Sanders, H. C. Gerritsen, “Fluorescence lifetime imaging, a new tool in confocal microscopy,” in Handbook of Biological Confocal Microscopy, J. Pawley, ed. (Plenum, New York, 1995), pp. 491–505.

D. W. Piston, D. R. Sandison, W. W. Webb, “Time-resolved fluorescence imaging and background rejection by two-photon excitation in laser scanning microscopy,” in Time-Resolved Laser Spectroscopy in Biochemistry III, J. R. Lakowicz, ed., Proc. SPIE1640, 379–389 (1992).

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