Abstract

We present a time-gated, optically sectioned, hyperspectral fluorescence lifetime imaging (FLIM) microscope incorporating a tunable supercontinuum excitation source extending into the UV. The system is capable of resolving the excitation spectrum, emission spectrum, and fluorescence decays in an optically sectioned image.

© 2007 Optical Society of America

Full Article  |  PDF Article

References

  • View by:
  • |
  • |

  1. D. K. Bird, K. W. Eliceiri, C. Fan, and J. White, Appl. Opt. 43, 5173 (2004).
    [CrossRef] [PubMed]
  2. P. De Beule, D. M. Owen, H. B. Manning, C. B. Talbot, J. Requejo-Isidro, C. Dunsby, J. McGinty, R. K. P. Benninger, D. S. Elson, I. Munro, M. J. Lever, P. Anand, M. A. A. Neil, and P. M. W. French, Microsc. Res. Tech. 70, 481 (2007).
    [CrossRef] [PubMed]
  3. D. S. Elson, N. Galletly, C. Talbot, J. Requejo-Isidro, J. McGinty, C. Dunsby, P. M. P. Lanigan, I. Munro, R. K. P. Benninger, P. De Beule, E. Auksorius, L. Hegyi, A. Sandison, A. Wallace, P. Soutter, M. A. A. Neil, J. Lever, G. W. Stamp, and P. M. W. French, in Reviews in Fluorescence 2006, C.D.Geddes and J.R.Lakowicz, eds. (Springer Science, 2006).
  4. R. Richards-Kortum and E. M. Sevick-Muraca, Annu. Rev. Phys. Chem. 47, 555 (1996).
    [CrossRef] [PubMed]
  5. H. B. Manning, D. M. Owen, E. Auksorius, P. A. A. De Beule, S. Oddos, C. B. Talbot, C. Dunsby, I. Munro, A. I. Magee, M. A. A. Neil, and P. M. W. French, in Proc. SPIE 6630, 663019 (2007).
    [CrossRef]
  6. E. A. Jares-Erijman and T. M. Jovin, Curr. Opin. Chem. Biol. 10, 409 (2006).
    [CrossRef] [PubMed]
  7. N. Deguil, E. Mottey, F. Salin, P. Legros, and D. Choquet, Microsc. Res. Tech. 63, 23 (2004).
    [CrossRef]
  8. G. McConnell, Opt. Express 12, 2844 (2004).
    [CrossRef] [PubMed]
  9. 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]
  10. A. Kudlinski, A. K. George, J. C. Knight, J. C. Travers, A. B. Rulkov, S. V. Popov, and J. R. Taylor, Opt. Express 14, 5715 (2006).
    [CrossRef] [PubMed]
  11. C. Sheppard and X. Mao, J. Mod. Opt. 35, 1169 (1988).
    [CrossRef]
  12. L. L. Nuffer, P. A. Medvick, H. P. Foote, and J. C. Solinsky, Cytometry 69A, 897 (2006).
    [CrossRef]

2007 (2)

H. B. Manning, D. M. Owen, E. Auksorius, P. A. A. De Beule, S. Oddos, C. B. Talbot, C. Dunsby, I. Munro, A. I. Magee, M. A. A. Neil, and P. M. W. French, in Proc. SPIE 6630, 663019 (2007).
[CrossRef]

P. De Beule, D. M. Owen, H. B. Manning, C. B. Talbot, J. Requejo-Isidro, C. Dunsby, J. McGinty, R. K. P. Benninger, D. S. Elson, I. Munro, M. J. Lever, P. Anand, M. A. A. Neil, and P. M. W. French, Microsc. Res. Tech. 70, 481 (2007).
[CrossRef] [PubMed]

2006 (3)

L. L. Nuffer, P. A. Medvick, H. P. Foote, and J. C. Solinsky, Cytometry 69A, 897 (2006).
[CrossRef]

E. A. Jares-Erijman and T. M. Jovin, Curr. Opin. Chem. Biol. 10, 409 (2006).
[CrossRef] [PubMed]

A. Kudlinski, A. K. George, J. C. Knight, J. C. Travers, A. B. Rulkov, S. V. Popov, and J. R. Taylor, Opt. Express 14, 5715 (2006).
[CrossRef] [PubMed]

2004 (4)

N. Deguil, E. Mottey, F. Salin, P. Legros, and D. Choquet, Microsc. Res. Tech. 63, 23 (2004).
[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]

G. McConnell, Opt. Express 12, 2844 (2004).
[CrossRef] [PubMed]

D. K. Bird, K. W. Eliceiri, C. Fan, and J. White, Appl. Opt. 43, 5173 (2004).
[CrossRef] [PubMed]

1996 (1)

R. Richards-Kortum and E. M. Sevick-Muraca, Annu. Rev. Phys. Chem. 47, 555 (1996).
[CrossRef] [PubMed]

1988 (1)

C. Sheppard and X. Mao, J. Mod. Opt. 35, 1169 (1988).
[CrossRef]

Annu. Rev. Phys. Chem. (1)

R. Richards-Kortum and E. M. Sevick-Muraca, Annu. Rev. Phys. Chem. 47, 555 (1996).
[CrossRef] [PubMed]

Appl. Opt. (1)

Curr. Opin. Chem. Biol. (1)

E. A. Jares-Erijman and T. M. Jovin, Curr. Opin. Chem. Biol. 10, 409 (2006).
[CrossRef] [PubMed]

Cytometry (1)

L. L. Nuffer, P. A. Medvick, H. P. Foote, and J. C. Solinsky, Cytometry 69A, 897 (2006).
[CrossRef]

J. Mod. Opt. (1)

C. Sheppard and X. Mao, J. Mod. Opt. 35, 1169 (1988).
[CrossRef]

J. Phys. D (1)

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]

Microsc. Res. Tech. (2)

N. Deguil, E. Mottey, F. Salin, P. Legros, and D. Choquet, Microsc. Res. Tech. 63, 23 (2004).
[CrossRef]

P. De Beule, D. M. Owen, H. B. Manning, C. B. Talbot, J. Requejo-Isidro, C. Dunsby, J. McGinty, R. K. P. Benninger, D. S. Elson, I. Munro, M. J. Lever, P. Anand, M. A. A. Neil, and P. M. W. French, Microsc. Res. Tech. 70, 481 (2007).
[CrossRef] [PubMed]

Opt. Express (2)

Proc. SPIE (1)

H. B. Manning, D. M. Owen, E. Auksorius, P. A. A. De Beule, S. Oddos, C. B. Talbot, C. Dunsby, I. Munro, A. I. Magee, M. A. A. Neil, and P. M. W. French, in Proc. SPIE 6630, 663019 (2007).
[CrossRef]

Other (1)

D. S. Elson, N. Galletly, C. Talbot, J. Requejo-Isidro, J. McGinty, C. Dunsby, P. M. P. Lanigan, I. Munro, R. K. P. Benninger, P. De Beule, E. Auksorius, L. Hegyi, A. Sandison, A. Wallace, P. Soutter, M. A. A. Neil, J. Lever, G. W. Stamp, and P. M. W. French, in Reviews in Fluorescence 2006, C.D.Geddes and J.R.Lakowicz, eds. (Springer Science, 2006).

Cited By

OSA participates in CrossRef's Cited-By Linking service. Citing articles from OSA journals and other participating publishers are listed here.

Alert me when this article is cited.


Figures (3)

Fig. 1
Fig. 1

Schematic of excitation-resolved hyperspectral FLIM microscope with spectral power density (SPD) of the continuum source extending into the UV.

Fig. 2
Fig. 2

Fluorescence data from unstained rat tail excited at 360 nm . (a) Intensity image showing, A, hair follicles; B, dermis; and, C, collagen bundles. (b) Selected emission spectra of these regions. (c) Mean emission wavelength image. (d), (e) FLIM images integrated over the two different emission spectral windows indicated.

Fig. 3
Fig. 3

Excitation-emission-lifetime (EEL) resolved fluorescence imaging of stained Convallaria. (a) Integrated fluorescence intensity image. (b) Mean emission wavelength image. (c) FLIM image for excitation at 470 nm . (d) EEM corresponding to regions indicated in (a). (e) Selected fluorescence decay profiles corresponding to regions α, β, and γ, respectively, indicated in (d) (normalized and scaled for clarity). (f) Plot of mean fluorescence lifetime as a function of emission and excitation wavelength.

Metrics