Abstract

Biophysical imaging tools exploit several properties of fluorescence to map cellular biochemistry. However, the engineering of a cost-effective and user-friendly detection system for sensing the diverse properties of fluorescence is a difficult challenge. Here, we present a novel architecture for a spectrograph that permits integrated characterization of excitation, emission and fluorescence anisotropy spectra in a quantitative and efficient manner. This sensing platform achieves excellent versatility of use at comparatively low costs. We demonstrate the novel optical design with example images of plant cells and of mammalian cells expressing fluorescent proteins undergoing energy transfer.

©2011 Optical Society of America

Full Article  |  PDF Article
OSA Recommended Articles
Confocal fluorescence spectroscopy and anisotropy imaging system

Chad E. Bigelow, David L. Conover, and Thomas H. Foster
Opt. Lett. 28(9) 695-697 (2003)

Imaging of protein cluster sizes by means of confocal time-gated fluorescence anisotropy microscopy

Arjen N. Bader, Erik G. Hofman, Paul M. P. van Bergen en Henegouwen, and Hans C. Gerritsen
Opt. Express 15(11) 6934-6945 (2007)

Time-resolved fluorescence anisotropy imaging applied to live cells

Klaus Suhling, Jan Siegel, Peter M. P. Lanigan, Sandrine Lévêque-Fort, Stephen E. D. Webb, David Phillips, Daniel M. Davis, and Paul M. W. French
Opt. Lett. 29(6) 584-586 (2004)

More Recommended Articles

References

  • View by:
  • Article Order
  • |
  • Year
  • |
  • Author
  • |
  • Publication
  1. R. A. Neher, M. Mitkovski, F. Kirchhoff, E. Neher, F. J. Theis, and A. Zeug, “Blind source separation techniques for the decomposition of multiply labeled fluorescence images,” Biophys. J. 96(9), 3791–3800 (2009).
    [Crossref] [PubMed]
  2. J. A. Palero, H. S. de Bruijn, A. van der Ploeg-van den Heuvel, H. J. C. M. Sterenborg, and H. C. Gerritsen, “In vivo nonlinear spectral imaging in mouse skin,” Opt. Express 14(10), 4395–4402 (2006).
    [Crossref] [PubMed]
  3. P. L. T. M. Frederix, M. A. H. Asselbergs, W. G. J. H. van Sark, D. J. van den Heuvel, W. Hamelink, E. L. de Beer, and H. C. Gerritsen, “High sensitivity spectrograph for use in fluorescence microscopy,” Appl. Spectrosc. 55(8), 1005–1012 (2001).
    [Crossref]
  4. T. S. Forde and Q. S. Hanley, “Spectrally resolved frequency domain analysis of multi-fluorophore systems undergoing energy transfer,” Appl. Spectrosc. 60(12), 1442–1452 (2006).
    [Crossref]
  5. A. Esposito, M. Gralle, M. A. C. Dani, D. Lange, and F. S. Wouters, “pHlameleons: a family of FRET-based protein sensors for quantitative pH imaging,” Biochemistry 47(49), 13115–13126 (2008).
    [Crossref] [PubMed]
  6. A. Esposito, T. Tiffert, J. M. Mauritz, S. Schlachter, L. H. Bannister, C. F. Kaminski, and V. L. Lew, “FRET imaging of hemoglobin concentration in Plasmodium falciparum-infected red cells,” PLoS ONE 3(11), e3780 (2008).
    [Crossref] [PubMed]
  7. A. N. Bader, E. G. Hofman, P. M. van Bergen En Henegouwen, and H. C. Gerritsen, “Imaging of protein cluster sizes by means of confocal time-gated fluorescence anisotropy microscopy,” Opt. Express 15(11), 6934–6945 (2007).
    [Crossref] [PubMed]
  8. G. J. Kremers, E. B. van Munster, J. Goedhart, and T. W. Gadella., “Quantitative lifetime unmixing of multiexponentially decaying fluorophores using single-frequency fluorescence lifetime imaging microscopy,” Biophys. J. 95(1), 378–389 (2008).
    [Crossref] [PubMed]
  9. S. Schlachter, S. Schwedler, A. Esposito, G. S. Kaminski Schierle, G. D. Moggridge, and C. F. Kaminski, “A method to unmix multiple fluorophores in microscopy images with minimal a priori information,” Opt. Express 17(25), 22747–22760 (2009).
    [Crossref]
  10. D. S. Lidke, P. Nagy, B. G. Barisas, R. Heintzmann, J. N. Post, K. A. Lidke, A. H. Clayton, D. J. Arndt-Jovin, and T. M. Jovin, “Imaging molecular interactions in cells by dynamic and static fluorescence anisotropy (rFLIM and emFRET),” Biochem. Soc. Trans. 31(5), 1020–1027 (2003).
    [Crossref] [PubMed]
  11. K. A. Lidke, B. Rieger, D. S. Lidke, and T. M. Jovin, “The role of photon statistics in fluorescence anisotropy imaging,” IEEE Trans. Image Process. 14(9), 1237–1245 (2005).
    [Crossref] [PubMed]
  12. A. H. A. Clayton, Q. S. Hanley, D. J. Arndt-Jovin, V. Subramaniam, and T. M. Jovin, “Dynamic fluorescence anisotropy imaging microscopy in the frequency domain (rFLIM),” Biophys. J. 83(3), 1631–1649 (2002).
    [Crossref] [PubMed]
  13. D. A. Bachovchin, S. J. Brown, H. Rosen, and B. F. Cravatt, “Identification of selective inhibitors of uncharacterized enzymes by high-throughput screening with fluorescent activity-based probes,” Nat. Biotechnol. 27(4), 387–394 (2009).
    [Crossref] [PubMed]
  14. D. R. Matthews, L. M. Carlin, E. Ofo, P. R. Barber, B. Vojnovic, M. Irving, T. Ng, and S. M. Ameer-Beg, “Time-lapse FRET microscopy using fluorescence anisotropy,” J. Microsc. 237(1), 51–62 (2010).
    [Crossref] [PubMed]
  15. F. T. S. Chan, C. F. Kaminski, and G. S. Kaminski Schierle, “HomoFRET fluorescence anisotropy imaging as a tool to study molecular self-assembly in live cells,” ChemPhysChem (2011), doi:.
    [Crossref] [PubMed]
  16. B. Valeur and G. Weber, “Resolution of the fluorescence excitation spectrum of indole into the 1La and 1Lb excitation bands,” Photochem. Photobiol. 25(5), 441–444 (1977).
    [Crossref] [PubMed]
  17. P. Blandin, S. Lévêque-Fort, S. Lécart, J. C. Cossec, M.-C. Potier, Z. Lenkei, F. Druon, and P. Georges, “Time-gated total internal reflection fluorescence microscopy with a supercontinuum excitation source,” Appl. Opt. 48(3), 553–559 (2009).
    [Crossref] [PubMed]
  18. J. H. Frank, A. D. Elder, J. Swartling, A. R. Venkitaraman, A. D. Jeyasekharan, and C. F. Kaminski, “A white light confocal microscope for spectrally resolved multidimensional imaging,” J. Microsc. 227(3), 203–215 (2007).
    [Crossref] [PubMed]
  19. J. Y. Ye, C. J. Divin, J. R. Baker, and T. B. Norris, “Whole spectrum fluorescence detection with ultrafast white light excitation,” Opt. Express 15(16), 10439–10445 (2007).
    [Crossref] [PubMed]
  20. C. Dunsby, P. M. P. Lanigan, J. McGinty, D. S. Elson, J. Requejo-Isidro, I. Munro, N. Galletly, F. McCann, B. Treanor, B. Önfelt, D. M. Davis, M A A. Neil, and P. M. W. French, “An electronically tunable ultrafast laser source applied to fluorescence imaging and fluorescence lifetime imaging microscopy,” J. Phys. D Appl. Phys. 37(23), 3296–3303 (2004).
    [Crossref]
  21. G. McConnell, “Confocal laser scanning fluorescence microscopy with a visible continuum source,” Opt. Express 12(13), 2844–2850 (2004).
    [Crossref] [PubMed]
  22. M. A. Digman, V. R. Caiolfa, M. Zamai, and E. Gratton, “The phasor approach to fluorescence lifetime imaging analysis,” Biophys. J. 94(2), L14–L16 (2008).
    [Crossref]
  23. Q. S. Hanley and A. H. Clayton, “AB-plot assisted determination of fluorophore mixtures in a fluorescence lifetime microscope using spectra or quenchers,” J. Microsc. 218(1), 62–67 (2005).
    [Crossref] [PubMed]
  24. S. Schlachter, A. D. Elder, A. Esposito, G. S. Kaminski, J. H. Frank, L. K. van Geest, and C. F. Kaminski, “mhFLIM: resolution of heterogeneous fluorescence decays in widefield lifetime microscopy,” Opt. Express 17(3), 1557–1570 (2009).
    [Crossref] [PubMed]
  25. M. A. Rizzo and D. W. Piston, “High-contrast imaging of fluorescent protein FRET by fluorescence polarization microscopy,” Biophys. J. 88(2), L14–L16 (2005).
    [Crossref]
  26. T. J. van Ham, A. Esposito, J. R. Kumita, S. T. D. Hsu, G. S. Kaminski Schierle, C. F. Kaminski, C. M. Dobson, E. A. A. Nollen, and C. W. Bertoncini, “Towards multiparametric fluorescent imaging of amyloid formation: studies of a YFP model of alpha-synuclein aggregation,” J. Mol. Biol. 395(3), 627–642 (2010).
    [Crossref]
  27. H. B. Manning, G. T. Kennedy, D. M. Owen, D. M. Grant, A. I. Magee, M. A. Neil, Y. Itoh, C. Dunsby, and P. M. French, “A compact, multidimensional spectrofluorometer exploiting supercontinuum generation,” J. Biophoton. 1(6), 494 (2008).
    [Crossref]
  28. H. C. Gerritsen, A. V. Agronskaia, A. N. Bader, and A. Esposito, “Time Domain FLIM: theory, Instrumentation and data analysis,” in FRET & FLIM Imaging Techniques, T. W. Gadella, ed. (Elsevier, Amsterdam, The Netherlands, 2009).
  29. A. Esposito, H. C. Gerritsen, T. Oggier, F. Lustenberger, and F. S. Wouters, “Innovating lifetime microscopy: a compact and simple tool for life sciences, screening, and diagnostics,” J. Biomed. Opt. 11(3), 034016 (2006).
    [Crossref]
  30. L. Pancheri and D. Stoppa, “A SPAD-based Pixel Linear Array for High-Speed Time-Gated Fluorescence Lifetime Imaging,” 2009 Proc. of Esscirc, 429–432 (2009).
  31. D. M. Grant, W. Zhang, E. J. McGhee, T. D. Bunney, C. B. Talbot, S. Kumar, I. Munro, C. Dunsby, M. A. Neil, M. Katan, and P. M. French, “Multiplexed FRET to image multiple signaling events in live cells,” Biophys. J. 95(10), L69–L71 (2008).
    [Crossref] [PubMed]
  32. A. D. Jeyasekharan, N. Ayoub, R. Mahen, J. Ries, A. Esposito, E. Rajendra, H. Hattori, R. P. Kulkarni, and A. R. Venkitaraman, “DNA damage regulates the mobility of Brca2 within the nucleoplasm of living cells,” Proc. Natl. Acad. Sci. U.S.A. 107(50), 21937–21942 (2010).
    [Crossref] [PubMed]

2011 (1)

F. T. S. Chan, C. F. Kaminski, and G. S. Kaminski Schierle, “HomoFRET fluorescence anisotropy imaging as a tool to study molecular self-assembly in live cells,” ChemPhysChem (2011), doi:.
[Crossref] [PubMed]

2010 (3)

T. J. van Ham, A. Esposito, J. R. Kumita, S. T. D. Hsu, G. S. Kaminski Schierle, C. F. Kaminski, C. M. Dobson, E. A. A. Nollen, and C. W. Bertoncini, “Towards multiparametric fluorescent imaging of amyloid formation: studies of a YFP model of alpha-synuclein aggregation,” J. Mol. Biol. 395(3), 627–642 (2010).
[Crossref]

D. R. Matthews, L. M. Carlin, E. Ofo, P. R. Barber, B. Vojnovic, M. Irving, T. Ng, and S. M. Ameer-Beg, “Time-lapse FRET microscopy using fluorescence anisotropy,” J. Microsc. 237(1), 51–62 (2010).
[Crossref] [PubMed]

A. D. Jeyasekharan, N. Ayoub, R. Mahen, J. Ries, A. Esposito, E. Rajendra, H. Hattori, R. P. Kulkarni, and A. R. Venkitaraman, “DNA damage regulates the mobility of Brca2 within the nucleoplasm of living cells,” Proc. Natl. Acad. Sci. U.S.A. 107(50), 21937–21942 (2010).
[Crossref] [PubMed]

2009 (5)

2008 (6)

A. Esposito, M. Gralle, M. A. C. Dani, D. Lange, and F. S. Wouters, “pHlameleons: a family of FRET-based protein sensors for quantitative pH imaging,” Biochemistry 47(49), 13115–13126 (2008).
[Crossref] [PubMed]

A. Esposito, T. Tiffert, J. M. Mauritz, S. Schlachter, L. H. Bannister, C. F. Kaminski, and V. L. Lew, “FRET imaging of hemoglobin concentration in Plasmodium falciparum-infected red cells,” PLoS ONE 3(11), e3780 (2008).
[Crossref] [PubMed]

G. J. Kremers, E. B. van Munster, J. Goedhart, and T. W. Gadella., “Quantitative lifetime unmixing of multiexponentially decaying fluorophores using single-frequency fluorescence lifetime imaging microscopy,” Biophys. J. 95(1), 378–389 (2008).
[Crossref] [PubMed]

D. M. Grant, W. Zhang, E. J. McGhee, T. D. Bunney, C. B. Talbot, S. Kumar, I. Munro, C. Dunsby, M. A. Neil, M. Katan, and P. M. French, “Multiplexed FRET to image multiple signaling events in live cells,” Biophys. J. 95(10), L69–L71 (2008).
[Crossref] [PubMed]

H. B. Manning, G. T. Kennedy, D. M. Owen, D. M. Grant, A. I. Magee, M. A. Neil, Y. Itoh, C. Dunsby, and P. M. French, “A compact, multidimensional spectrofluorometer exploiting supercontinuum generation,” J. Biophoton. 1(6), 494 (2008).
[Crossref]

M. A. Digman, V. R. Caiolfa, M. Zamai, and E. Gratton, “The phasor approach to fluorescence lifetime imaging analysis,” Biophys. J. 94(2), L14–L16 (2008).
[Crossref]

2007 (3)

2006 (3)

2005 (3)

Q. S. Hanley and A. H. Clayton, “AB-plot assisted determination of fluorophore mixtures in a fluorescence lifetime microscope using spectra or quenchers,” J. Microsc. 218(1), 62–67 (2005).
[Crossref] [PubMed]

M. A. Rizzo and D. W. Piston, “High-contrast imaging of fluorescent protein FRET by fluorescence polarization microscopy,” Biophys. J. 88(2), L14–L16 (2005).
[Crossref]

K. A. Lidke, B. Rieger, D. S. Lidke, and T. M. Jovin, “The role of photon statistics in fluorescence anisotropy imaging,” IEEE Trans. Image Process. 14(9), 1237–1245 (2005).
[Crossref] [PubMed]

2004 (2)

C. Dunsby, P. M. P. Lanigan, J. McGinty, D. S. Elson, J. Requejo-Isidro, I. Munro, N. Galletly, F. McCann, B. Treanor, B. Önfelt, D. M. Davis, M A A. Neil, and P. M. W. French, “An electronically tunable ultrafast laser source applied to fluorescence imaging and fluorescence lifetime imaging microscopy,” J. Phys. D Appl. Phys. 37(23), 3296–3303 (2004).
[Crossref]

G. McConnell, “Confocal laser scanning fluorescence microscopy with a visible continuum source,” Opt. Express 12(13), 2844–2850 (2004).
[Crossref] [PubMed]

2003 (1)

D. S. Lidke, P. Nagy, B. G. Barisas, R. Heintzmann, J. N. Post, K. A. Lidke, A. H. Clayton, D. J. Arndt-Jovin, and T. M. Jovin, “Imaging molecular interactions in cells by dynamic and static fluorescence anisotropy (rFLIM and emFRET),” Biochem. Soc. Trans. 31(5), 1020–1027 (2003).
[Crossref] [PubMed]

2002 (1)

A. H. A. Clayton, Q. S. Hanley, D. J. Arndt-Jovin, V. Subramaniam, and T. M. Jovin, “Dynamic fluorescence anisotropy imaging microscopy in the frequency domain (rFLIM),” Biophys. J. 83(3), 1631–1649 (2002).
[Crossref] [PubMed]

2001 (1)

1977 (1)

B. Valeur and G. Weber, “Resolution of the fluorescence excitation spectrum of indole into the 1La and 1Lb excitation bands,” Photochem. Photobiol. 25(5), 441–444 (1977).
[Crossref] [PubMed]

Ameer-Beg, S. M.

D. R. Matthews, L. M. Carlin, E. Ofo, P. R. Barber, B. Vojnovic, M. Irving, T. Ng, and S. M. Ameer-Beg, “Time-lapse FRET microscopy using fluorescence anisotropy,” J. Microsc. 237(1), 51–62 (2010).
[Crossref] [PubMed]

Arndt-Jovin, D. J.

D. S. Lidke, P. Nagy, B. G. Barisas, R. Heintzmann, J. N. Post, K. A. Lidke, A. H. Clayton, D. J. Arndt-Jovin, and T. M. Jovin, “Imaging molecular interactions in cells by dynamic and static fluorescence anisotropy (rFLIM and emFRET),” Biochem. Soc. Trans. 31(5), 1020–1027 (2003).
[Crossref] [PubMed]

A. H. A. Clayton, Q. S. Hanley, D. J. Arndt-Jovin, V. Subramaniam, and T. M. Jovin, “Dynamic fluorescence anisotropy imaging microscopy in the frequency domain (rFLIM),” Biophys. J. 83(3), 1631–1649 (2002).
[Crossref] [PubMed]

Asselbergs, M. A. H.

Ayoub, N.

A. D. Jeyasekharan, N. Ayoub, R. Mahen, J. Ries, A. Esposito, E. Rajendra, H. Hattori, R. P. Kulkarni, and A. R. Venkitaraman, “DNA damage regulates the mobility of Brca2 within the nucleoplasm of living cells,” Proc. Natl. Acad. Sci. U.S.A. 107(50), 21937–21942 (2010).
[Crossref] [PubMed]

Bachovchin, D. A.

D. A. Bachovchin, S. J. Brown, H. Rosen, and B. F. Cravatt, “Identification of selective inhibitors of uncharacterized enzymes by high-throughput screening with fluorescent activity-based probes,” Nat. Biotechnol. 27(4), 387–394 (2009).
[Crossref] [PubMed]

Bader, A. N.

Baker, J. R.

Bannister, L. H.

A. Esposito, T. Tiffert, J. M. Mauritz, S. Schlachter, L. H. Bannister, C. F. Kaminski, and V. L. Lew, “FRET imaging of hemoglobin concentration in Plasmodium falciparum-infected red cells,” PLoS ONE 3(11), e3780 (2008).
[Crossref] [PubMed]

Barber, P. R.

D. R. Matthews, L. M. Carlin, E. Ofo, P. R. Barber, B. Vojnovic, M. Irving, T. Ng, and S. M. Ameer-Beg, “Time-lapse FRET microscopy using fluorescence anisotropy,” J. Microsc. 237(1), 51–62 (2010).
[Crossref] [PubMed]

Barisas, B. G.

D. S. Lidke, P. Nagy, B. G. Barisas, R. Heintzmann, J. N. Post, K. A. Lidke, A. H. Clayton, D. J. Arndt-Jovin, and T. M. Jovin, “Imaging molecular interactions in cells by dynamic and static fluorescence anisotropy (rFLIM and emFRET),” Biochem. Soc. Trans. 31(5), 1020–1027 (2003).
[Crossref] [PubMed]

Bertoncini, C. W.

T. J. van Ham, A. Esposito, J. R. Kumita, S. T. D. Hsu, G. S. Kaminski Schierle, C. F. Kaminski, C. M. Dobson, E. A. A. Nollen, and C. W. Bertoncini, “Towards multiparametric fluorescent imaging of amyloid formation: studies of a YFP model of alpha-synuclein aggregation,” J. Mol. Biol. 395(3), 627–642 (2010).
[Crossref]

Blandin, P.

Brown, S. J.

D. A. Bachovchin, S. J. Brown, H. Rosen, and B. F. Cravatt, “Identification of selective inhibitors of uncharacterized enzymes by high-throughput screening with fluorescent activity-based probes,” Nat. Biotechnol. 27(4), 387–394 (2009).
[Crossref] [PubMed]

Bunney, T. D.

D. M. Grant, W. Zhang, E. J. McGhee, T. D. Bunney, C. B. Talbot, S. Kumar, I. Munro, C. Dunsby, M. A. Neil, M. Katan, and P. M. French, “Multiplexed FRET to image multiple signaling events in live cells,” Biophys. J. 95(10), L69–L71 (2008).
[Crossref] [PubMed]

Caiolfa, V. R.

M. A. Digman, V. R. Caiolfa, M. Zamai, and E. Gratton, “The phasor approach to fluorescence lifetime imaging analysis,” Biophys. J. 94(2), L14–L16 (2008).
[Crossref]

Carlin, L. M.

D. R. Matthews, L. M. Carlin, E. Ofo, P. R. Barber, B. Vojnovic, M. Irving, T. Ng, and S. M. Ameer-Beg, “Time-lapse FRET microscopy using fluorescence anisotropy,” J. Microsc. 237(1), 51–62 (2010).
[Crossref] [PubMed]

Chan, F. T. S.

F. T. S. Chan, C. F. Kaminski, and G. S. Kaminski Schierle, “HomoFRET fluorescence anisotropy imaging as a tool to study molecular self-assembly in live cells,” ChemPhysChem (2011), doi:.
[Crossref] [PubMed]

Clayton, A. H.

Q. S. Hanley and A. H. Clayton, “AB-plot assisted determination of fluorophore mixtures in a fluorescence lifetime microscope using spectra or quenchers,” J. Microsc. 218(1), 62–67 (2005).
[Crossref] [PubMed]

D. S. Lidke, P. Nagy, B. G. Barisas, R. Heintzmann, J. N. Post, K. A. Lidke, A. H. Clayton, D. J. Arndt-Jovin, and T. M. Jovin, “Imaging molecular interactions in cells by dynamic and static fluorescence anisotropy (rFLIM and emFRET),” Biochem. Soc. Trans. 31(5), 1020–1027 (2003).
[Crossref] [PubMed]

Clayton, A. H. A.

A. H. A. Clayton, Q. S. Hanley, D. J. Arndt-Jovin, V. Subramaniam, and T. M. Jovin, “Dynamic fluorescence anisotropy imaging microscopy in the frequency domain (rFLIM),” Biophys. J. 83(3), 1631–1649 (2002).
[Crossref] [PubMed]

Cossec, J. C.

Cravatt, B. F.

D. A. Bachovchin, S. J. Brown, H. Rosen, and B. F. Cravatt, “Identification of selective inhibitors of uncharacterized enzymes by high-throughput screening with fluorescent activity-based probes,” Nat. Biotechnol. 27(4), 387–394 (2009).
[Crossref] [PubMed]

Dani, M. A. C.

A. Esposito, M. Gralle, M. A. C. Dani, D. Lange, and F. S. Wouters, “pHlameleons: a family of FRET-based protein sensors for quantitative pH imaging,” Biochemistry 47(49), 13115–13126 (2008).
[Crossref] [PubMed]

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. Önfelt, D. M. Davis, M A A. Neil, and P. M. W. French, “An electronically tunable ultrafast laser source applied to fluorescence imaging and fluorescence lifetime imaging microscopy,” J. Phys. D Appl. Phys. 37(23), 3296–3303 (2004).
[Crossref]

de Beer, E. L.

de Bruijn, H. S.

Digman, M. A.

M. A. Digman, V. R. Caiolfa, M. Zamai, and E. Gratton, “The phasor approach to fluorescence lifetime imaging analysis,” Biophys. J. 94(2), L14–L16 (2008).
[Crossref]

Divin, C. J.

Dobson, C. M.

T. J. van Ham, A. Esposito, J. R. Kumita, S. T. D. Hsu, G. S. Kaminski Schierle, C. F. Kaminski, C. M. Dobson, E. A. A. Nollen, and C. W. Bertoncini, “Towards multiparametric fluorescent imaging of amyloid formation: studies of a YFP model of alpha-synuclein aggregation,” J. Mol. Biol. 395(3), 627–642 (2010).
[Crossref]

Druon, F.

Dunsby, C.

D. M. Grant, W. Zhang, E. J. McGhee, T. D. Bunney, C. B. Talbot, S. Kumar, I. Munro, C. Dunsby, M. A. Neil, M. Katan, and P. M. French, “Multiplexed FRET to image multiple signaling events in live cells,” Biophys. J. 95(10), L69–L71 (2008).
[Crossref] [PubMed]

H. B. Manning, G. T. Kennedy, D. M. Owen, D. M. Grant, A. I. Magee, M. A. Neil, Y. Itoh, C. Dunsby, and P. M. French, “A compact, multidimensional spectrofluorometer exploiting supercontinuum generation,” J. Biophoton. 1(6), 494 (2008).
[Crossref]

C. Dunsby, P. M. P. Lanigan, J. McGinty, D. S. Elson, J. Requejo-Isidro, I. Munro, N. Galletly, F. McCann, B. Treanor, B. Önfelt, D. M. Davis, M A A. Neil, and P. M. W. French, “An electronically tunable ultrafast laser source applied to fluorescence imaging and fluorescence lifetime imaging microscopy,” J. Phys. D Appl. Phys. 37(23), 3296–3303 (2004).
[Crossref]

Elder, A. D.

S. Schlachter, A. D. Elder, A. Esposito, G. S. Kaminski, J. H. Frank, L. K. van Geest, and C. F. Kaminski, “mhFLIM: resolution of heterogeneous fluorescence decays in widefield lifetime microscopy,” Opt. Express 17(3), 1557–1570 (2009).
[Crossref] [PubMed]

J. H. Frank, A. D. Elder, J. Swartling, A. R. Venkitaraman, A. D. Jeyasekharan, and C. F. Kaminski, “A white light confocal microscope for spectrally resolved multidimensional imaging,” J. Microsc. 227(3), 203–215 (2007).
[Crossref] [PubMed]

Elson, D. S.

C. Dunsby, P. M. P. Lanigan, J. McGinty, D. S. Elson, J. Requejo-Isidro, I. Munro, N. Galletly, F. McCann, B. Treanor, B. Önfelt, D. M. Davis, M A A. Neil, and P. M. W. French, “An electronically tunable ultrafast laser source applied to fluorescence imaging and fluorescence lifetime imaging microscopy,” J. Phys. D Appl. Phys. 37(23), 3296–3303 (2004).
[Crossref]

Esposito, A.

T. J. van Ham, A. Esposito, J. R. Kumita, S. T. D. Hsu, G. S. Kaminski Schierle, C. F. Kaminski, C. M. Dobson, E. A. A. Nollen, and C. W. Bertoncini, “Towards multiparametric fluorescent imaging of amyloid formation: studies of a YFP model of alpha-synuclein aggregation,” J. Mol. Biol. 395(3), 627–642 (2010).
[Crossref]

A. D. Jeyasekharan, N. Ayoub, R. Mahen, J. Ries, A. Esposito, E. Rajendra, H. Hattori, R. P. Kulkarni, and A. R. Venkitaraman, “DNA damage regulates the mobility of Brca2 within the nucleoplasm of living cells,” Proc. Natl. Acad. Sci. U.S.A. 107(50), 21937–21942 (2010).
[Crossref] [PubMed]

S. Schlachter, A. D. Elder, A. Esposito, G. S. Kaminski, J. H. Frank, L. K. van Geest, and C. F. Kaminski, “mhFLIM: resolution of heterogeneous fluorescence decays in widefield lifetime microscopy,” Opt. Express 17(3), 1557–1570 (2009).
[Crossref] [PubMed]

S. Schlachter, S. Schwedler, A. Esposito, G. S. Kaminski Schierle, G. D. Moggridge, and C. F. Kaminski, “A method to unmix multiple fluorophores in microscopy images with minimal a priori information,” Opt. Express 17(25), 22747–22760 (2009).
[Crossref]

A. Esposito, M. Gralle, M. A. C. Dani, D. Lange, and F. S. Wouters, “pHlameleons: a family of FRET-based protein sensors for quantitative pH imaging,” Biochemistry 47(49), 13115–13126 (2008).
[Crossref] [PubMed]

A. Esposito, T. Tiffert, J. M. Mauritz, S. Schlachter, L. H. Bannister, C. F. Kaminski, and V. L. Lew, “FRET imaging of hemoglobin concentration in Plasmodium falciparum-infected red cells,” PLoS ONE 3(11), e3780 (2008).
[Crossref] [PubMed]

A. Esposito, H. C. Gerritsen, T. Oggier, F. Lustenberger, and F. S. Wouters, “Innovating lifetime microscopy: a compact and simple tool for life sciences, screening, and diagnostics,” J. Biomed. Opt. 11(3), 034016 (2006).
[Crossref]

Forde, T. S.

Frank, J. H.

S. Schlachter, A. D. Elder, A. Esposito, G. S. Kaminski, J. H. Frank, L. K. van Geest, and C. F. Kaminski, “mhFLIM: resolution of heterogeneous fluorescence decays in widefield lifetime microscopy,” Opt. Express 17(3), 1557–1570 (2009).
[Crossref] [PubMed]

J. H. Frank, A. D. Elder, J. Swartling, A. R. Venkitaraman, A. D. Jeyasekharan, and C. F. Kaminski, “A white light confocal microscope for spectrally resolved multidimensional imaging,” J. Microsc. 227(3), 203–215 (2007).
[Crossref] [PubMed]

Frederix, P. L. T. M.

French, P. M.

D. M. Grant, W. Zhang, E. J. McGhee, T. D. Bunney, C. B. Talbot, S. Kumar, I. Munro, C. Dunsby, M. A. Neil, M. Katan, and P. M. French, “Multiplexed FRET to image multiple signaling events in live cells,” Biophys. J. 95(10), L69–L71 (2008).
[Crossref] [PubMed]

H. B. Manning, G. T. Kennedy, D. M. Owen, D. M. Grant, A. I. Magee, M. A. Neil, Y. Itoh, C. Dunsby, and P. M. French, “A compact, multidimensional spectrofluorometer exploiting supercontinuum generation,” J. Biophoton. 1(6), 494 (2008).
[Crossref]

French, P. M. W.

C. Dunsby, P. M. P. Lanigan, J. McGinty, D. S. Elson, J. Requejo-Isidro, I. Munro, N. Galletly, F. McCann, B. Treanor, B. Önfelt, D. M. Davis, M A A. Neil, and P. M. W. French, “An electronically tunable ultrafast laser source applied to fluorescence imaging and fluorescence lifetime imaging microscopy,” J. Phys. D Appl. Phys. 37(23), 3296–3303 (2004).
[Crossref]

Gadella, T. W.

G. J. Kremers, E. B. van Munster, J. Goedhart, and T. W. Gadella., “Quantitative lifetime unmixing of multiexponentially decaying fluorophores using single-frequency fluorescence lifetime imaging microscopy,” Biophys. J. 95(1), 378–389 (2008).
[Crossref] [PubMed]

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. Önfelt, D. M. Davis, M A A. Neil, and P. M. W. French, “An electronically tunable ultrafast laser source applied to fluorescence imaging and fluorescence lifetime imaging microscopy,” J. Phys. D Appl. Phys. 37(23), 3296–3303 (2004).
[Crossref]

Georges, P.

Gerritsen, H. C.

Goedhart, J.

G. J. Kremers, E. B. van Munster, J. Goedhart, and T. W. Gadella., “Quantitative lifetime unmixing of multiexponentially decaying fluorophores using single-frequency fluorescence lifetime imaging microscopy,” Biophys. J. 95(1), 378–389 (2008).
[Crossref] [PubMed]

Gralle, M.

A. Esposito, M. Gralle, M. A. C. Dani, D. Lange, and F. S. Wouters, “pHlameleons: a family of FRET-based protein sensors for quantitative pH imaging,” Biochemistry 47(49), 13115–13126 (2008).
[Crossref] [PubMed]

Grant, D. M.

H. B. Manning, G. T. Kennedy, D. M. Owen, D. M. Grant, A. I. Magee, M. A. Neil, Y. Itoh, C. Dunsby, and P. M. French, “A compact, multidimensional spectrofluorometer exploiting supercontinuum generation,” J. Biophoton. 1(6), 494 (2008).
[Crossref]

D. M. Grant, W. Zhang, E. J. McGhee, T. D. Bunney, C. B. Talbot, S. Kumar, I. Munro, C. Dunsby, M. A. Neil, M. Katan, and P. M. French, “Multiplexed FRET to image multiple signaling events in live cells,” Biophys. J. 95(10), L69–L71 (2008).
[Crossref] [PubMed]

Gratton, E.

M. A. Digman, V. R. Caiolfa, M. Zamai, and E. Gratton, “The phasor approach to fluorescence lifetime imaging analysis,” Biophys. J. 94(2), L14–L16 (2008).
[Crossref]

Hamelink, W.

Hanley, Q. S.

T. S. Forde and Q. S. Hanley, “Spectrally resolved frequency domain analysis of multi-fluorophore systems undergoing energy transfer,” Appl. Spectrosc. 60(12), 1442–1452 (2006).
[Crossref]

Q. S. Hanley and A. H. Clayton, “AB-plot assisted determination of fluorophore mixtures in a fluorescence lifetime microscope using spectra or quenchers,” J. Microsc. 218(1), 62–67 (2005).
[Crossref] [PubMed]

A. H. A. Clayton, Q. S. Hanley, D. J. Arndt-Jovin, V. Subramaniam, and T. M. Jovin, “Dynamic fluorescence anisotropy imaging microscopy in the frequency domain (rFLIM),” Biophys. J. 83(3), 1631–1649 (2002).
[Crossref] [PubMed]

Hattori, H.

A. D. Jeyasekharan, N. Ayoub, R. Mahen, J. Ries, A. Esposito, E. Rajendra, H. Hattori, R. P. Kulkarni, and A. R. Venkitaraman, “DNA damage regulates the mobility of Brca2 within the nucleoplasm of living cells,” Proc. Natl. Acad. Sci. U.S.A. 107(50), 21937–21942 (2010).
[Crossref] [PubMed]

Heintzmann, R.

D. S. Lidke, P. Nagy, B. G. Barisas, R. Heintzmann, J. N. Post, K. A. Lidke, A. H. Clayton, D. J. Arndt-Jovin, and T. M. Jovin, “Imaging molecular interactions in cells by dynamic and static fluorescence anisotropy (rFLIM and emFRET),” Biochem. Soc. Trans. 31(5), 1020–1027 (2003).
[Crossref] [PubMed]

Hofman, E. G.

Hsu, S. T. D.

T. J. van Ham, A. Esposito, J. R. Kumita, S. T. D. Hsu, G. S. Kaminski Schierle, C. F. Kaminski, C. M. Dobson, E. A. A. Nollen, and C. W. Bertoncini, “Towards multiparametric fluorescent imaging of amyloid formation: studies of a YFP model of alpha-synuclein aggregation,” J. Mol. Biol. 395(3), 627–642 (2010).
[Crossref]

Irving, M.

D. R. Matthews, L. M. Carlin, E. Ofo, P. R. Barber, B. Vojnovic, M. Irving, T. Ng, and S. M. Ameer-Beg, “Time-lapse FRET microscopy using fluorescence anisotropy,” J. Microsc. 237(1), 51–62 (2010).
[Crossref] [PubMed]

Itoh, Y.

H. B. Manning, G. T. Kennedy, D. M. Owen, D. M. Grant, A. I. Magee, M. A. Neil, Y. Itoh, C. Dunsby, and P. M. French, “A compact, multidimensional spectrofluorometer exploiting supercontinuum generation,” J. Biophoton. 1(6), 494 (2008).
[Crossref]

Jeyasekharan, A. D.

A. D. Jeyasekharan, N. Ayoub, R. Mahen, J. Ries, A. Esposito, E. Rajendra, H. Hattori, R. P. Kulkarni, and A. R. Venkitaraman, “DNA damage regulates the mobility of Brca2 within the nucleoplasm of living cells,” Proc. Natl. Acad. Sci. U.S.A. 107(50), 21937–21942 (2010).
[Crossref] [PubMed]

J. H. Frank, A. D. Elder, J. Swartling, A. R. Venkitaraman, A. D. Jeyasekharan, and C. F. Kaminski, “A white light confocal microscope for spectrally resolved multidimensional imaging,” J. Microsc. 227(3), 203–215 (2007).
[Crossref] [PubMed]

Jovin, T. M.

K. A. Lidke, B. Rieger, D. S. Lidke, and T. M. Jovin, “The role of photon statistics in fluorescence anisotropy imaging,” IEEE Trans. Image Process. 14(9), 1237–1245 (2005).
[Crossref] [PubMed]

D. S. Lidke, P. Nagy, B. G. Barisas, R. Heintzmann, J. N. Post, K. A. Lidke, A. H. Clayton, D. J. Arndt-Jovin, and T. M. Jovin, “Imaging molecular interactions in cells by dynamic and static fluorescence anisotropy (rFLIM and emFRET),” Biochem. Soc. Trans. 31(5), 1020–1027 (2003).
[Crossref] [PubMed]

A. H. A. Clayton, Q. S. Hanley, D. J. Arndt-Jovin, V. Subramaniam, and T. M. Jovin, “Dynamic fluorescence anisotropy imaging microscopy in the frequency domain (rFLIM),” Biophys. J. 83(3), 1631–1649 (2002).
[Crossref] [PubMed]

Kaminski, C. F.

F. T. S. Chan, C. F. Kaminski, and G. S. Kaminski Schierle, “HomoFRET fluorescence anisotropy imaging as a tool to study molecular self-assembly in live cells,” ChemPhysChem (2011), doi:.
[Crossref] [PubMed]

T. J. van Ham, A. Esposito, J. R. Kumita, S. T. D. Hsu, G. S. Kaminski Schierle, C. F. Kaminski, C. M. Dobson, E. A. A. Nollen, and C. W. Bertoncini, “Towards multiparametric fluorescent imaging of amyloid formation: studies of a YFP model of alpha-synuclein aggregation,” J. Mol. Biol. 395(3), 627–642 (2010).
[Crossref]

S. Schlachter, S. Schwedler, A. Esposito, G. S. Kaminski Schierle, G. D. Moggridge, and C. F. Kaminski, “A method to unmix multiple fluorophores in microscopy images with minimal a priori information,” Opt. Express 17(25), 22747–22760 (2009).
[Crossref]

S. Schlachter, A. D. Elder, A. Esposito, G. S. Kaminski, J. H. Frank, L. K. van Geest, and C. F. Kaminski, “mhFLIM: resolution of heterogeneous fluorescence decays in widefield lifetime microscopy,” Opt. Express 17(3), 1557–1570 (2009).
[Crossref] [PubMed]

A. Esposito, T. Tiffert, J. M. Mauritz, S. Schlachter, L. H. Bannister, C. F. Kaminski, and V. L. Lew, “FRET imaging of hemoglobin concentration in Plasmodium falciparum-infected red cells,” PLoS ONE 3(11), e3780 (2008).
[Crossref] [PubMed]

J. H. Frank, A. D. Elder, J. Swartling, A. R. Venkitaraman, A. D. Jeyasekharan, and C. F. Kaminski, “A white light confocal microscope for spectrally resolved multidimensional imaging,” J. Microsc. 227(3), 203–215 (2007).
[Crossref] [PubMed]

Kaminski, G. S.

Kaminski Schierle, G. S.

F. T. S. Chan, C. F. Kaminski, and G. S. Kaminski Schierle, “HomoFRET fluorescence anisotropy imaging as a tool to study molecular self-assembly in live cells,” ChemPhysChem (2011), doi:.
[Crossref] [PubMed]

T. J. van Ham, A. Esposito, J. R. Kumita, S. T. D. Hsu, G. S. Kaminski Schierle, C. F. Kaminski, C. M. Dobson, E. A. A. Nollen, and C. W. Bertoncini, “Towards multiparametric fluorescent imaging of amyloid formation: studies of a YFP model of alpha-synuclein aggregation,” J. Mol. Biol. 395(3), 627–642 (2010).
[Crossref]

S. Schlachter, S. Schwedler, A. Esposito, G. S. Kaminski Schierle, G. D. Moggridge, and C. F. Kaminski, “A method to unmix multiple fluorophores in microscopy images with minimal a priori information,” Opt. Express 17(25), 22747–22760 (2009).
[Crossref]

Katan, M.

D. M. Grant, W. Zhang, E. J. McGhee, T. D. Bunney, C. B. Talbot, S. Kumar, I. Munro, C. Dunsby, M. A. Neil, M. Katan, and P. M. French, “Multiplexed FRET to image multiple signaling events in live cells,” Biophys. J. 95(10), L69–L71 (2008).
[Crossref] [PubMed]

Kennedy, G. T.

H. B. Manning, G. T. Kennedy, D. M. Owen, D. M. Grant, A. I. Magee, M. A. Neil, Y. Itoh, C. Dunsby, and P. M. French, “A compact, multidimensional spectrofluorometer exploiting supercontinuum generation,” J. Biophoton. 1(6), 494 (2008).
[Crossref]

Kirchhoff, F.

R. A. Neher, M. Mitkovski, F. Kirchhoff, E. Neher, F. J. Theis, and A. Zeug, “Blind source separation techniques for the decomposition of multiply labeled fluorescence images,” Biophys. J. 96(9), 3791–3800 (2009).
[Crossref] [PubMed]

Kremers, G. J.

G. J. Kremers, E. B. van Munster, J. Goedhart, and T. W. Gadella., “Quantitative lifetime unmixing of multiexponentially decaying fluorophores using single-frequency fluorescence lifetime imaging microscopy,” Biophys. J. 95(1), 378–389 (2008).
[Crossref] [PubMed]

Kulkarni, R. P.

A. D. Jeyasekharan, N. Ayoub, R. Mahen, J. Ries, A. Esposito, E. Rajendra, H. Hattori, R. P. Kulkarni, and A. R. Venkitaraman, “DNA damage regulates the mobility of Brca2 within the nucleoplasm of living cells,” Proc. Natl. Acad. Sci. U.S.A. 107(50), 21937–21942 (2010).
[Crossref] [PubMed]

Kumar, S.

D. M. Grant, W. Zhang, E. J. McGhee, T. D. Bunney, C. B. Talbot, S. Kumar, I. Munro, C. Dunsby, M. A. Neil, M. Katan, and P. M. French, “Multiplexed FRET to image multiple signaling events in live cells,” Biophys. J. 95(10), L69–L71 (2008).
[Crossref] [PubMed]

Kumita, J. R.

T. J. van Ham, A. Esposito, J. R. Kumita, S. T. D. Hsu, G. S. Kaminski Schierle, C. F. Kaminski, C. M. Dobson, E. A. A. Nollen, and C. W. Bertoncini, “Towards multiparametric fluorescent imaging of amyloid formation: studies of a YFP model of alpha-synuclein aggregation,” J. Mol. Biol. 395(3), 627–642 (2010).
[Crossref]

Lange, D.

A. Esposito, M. Gralle, M. A. C. Dani, D. Lange, and F. S. Wouters, “pHlameleons: a family of FRET-based protein sensors for quantitative pH imaging,” Biochemistry 47(49), 13115–13126 (2008).
[Crossref] [PubMed]

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. Önfelt, D. M. Davis, M A A. Neil, and P. M. W. French, “An electronically tunable ultrafast laser source applied to fluorescence imaging and fluorescence lifetime imaging microscopy,” J. Phys. D Appl. Phys. 37(23), 3296–3303 (2004).
[Crossref]

Lécart, S.

Lenkei, Z.

Lévêque-Fort, S.

Lew, V. L.

A. Esposito, T. Tiffert, J. M. Mauritz, S. Schlachter, L. H. Bannister, C. F. Kaminski, and V. L. Lew, “FRET imaging of hemoglobin concentration in Plasmodium falciparum-infected red cells,” PLoS ONE 3(11), e3780 (2008).
[Crossref] [PubMed]

Lidke, D. S.

K. A. Lidke, B. Rieger, D. S. Lidke, and T. M. Jovin, “The role of photon statistics in fluorescence anisotropy imaging,” IEEE Trans. Image Process. 14(9), 1237–1245 (2005).
[Crossref] [PubMed]

D. S. Lidke, P. Nagy, B. G. Barisas, R. Heintzmann, J. N. Post, K. A. Lidke, A. H. Clayton, D. J. Arndt-Jovin, and T. M. Jovin, “Imaging molecular interactions in cells by dynamic and static fluorescence anisotropy (rFLIM and emFRET),” Biochem. Soc. Trans. 31(5), 1020–1027 (2003).
[Crossref] [PubMed]

Lidke, K. A.

K. A. Lidke, B. Rieger, D. S. Lidke, and T. M. Jovin, “The role of photon statistics in fluorescence anisotropy imaging,” IEEE Trans. Image Process. 14(9), 1237–1245 (2005).
[Crossref] [PubMed]

D. S. Lidke, P. Nagy, B. G. Barisas, R. Heintzmann, J. N. Post, K. A. Lidke, A. H. Clayton, D. J. Arndt-Jovin, and T. M. Jovin, “Imaging molecular interactions in cells by dynamic and static fluorescence anisotropy (rFLIM and emFRET),” Biochem. Soc. Trans. 31(5), 1020–1027 (2003).
[Crossref] [PubMed]

Lustenberger, F.

A. Esposito, H. C. Gerritsen, T. Oggier, F. Lustenberger, and F. S. Wouters, “Innovating lifetime microscopy: a compact and simple tool for life sciences, screening, and diagnostics,” J. Biomed. Opt. 11(3), 034016 (2006).
[Crossref]

Magee, A. I.

H. B. Manning, G. T. Kennedy, D. M. Owen, D. M. Grant, A. I. Magee, M. A. Neil, Y. Itoh, C. Dunsby, and P. M. French, “A compact, multidimensional spectrofluorometer exploiting supercontinuum generation,” J. Biophoton. 1(6), 494 (2008).
[Crossref]

Mahen, R.

A. D. Jeyasekharan, N. Ayoub, R. Mahen, J. Ries, A. Esposito, E. Rajendra, H. Hattori, R. P. Kulkarni, and A. R. Venkitaraman, “DNA damage regulates the mobility of Brca2 within the nucleoplasm of living cells,” Proc. Natl. Acad. Sci. U.S.A. 107(50), 21937–21942 (2010).
[Crossref] [PubMed]

Manning, H. B.

H. B. Manning, G. T. Kennedy, D. M. Owen, D. M. Grant, A. I. Magee, M. A. Neil, Y. Itoh, C. Dunsby, and P. M. French, “A compact, multidimensional spectrofluorometer exploiting supercontinuum generation,” J. Biophoton. 1(6), 494 (2008).
[Crossref]

Matthews, D. R.

D. R. Matthews, L. M. Carlin, E. Ofo, P. R. Barber, B. Vojnovic, M. Irving, T. Ng, and S. M. Ameer-Beg, “Time-lapse FRET microscopy using fluorescence anisotropy,” J. Microsc. 237(1), 51–62 (2010).
[Crossref] [PubMed]

Mauritz, J. M.

A. Esposito, T. Tiffert, J. M. Mauritz, S. Schlachter, L. H. Bannister, C. F. Kaminski, and V. L. Lew, “FRET imaging of hemoglobin concentration in Plasmodium falciparum-infected red cells,” PLoS ONE 3(11), e3780 (2008).
[Crossref] [PubMed]

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. Önfelt, D. M. Davis, M A A. Neil, and P. M. W. French, “An electronically tunable ultrafast laser source applied to fluorescence imaging and fluorescence lifetime imaging microscopy,” J. Phys. D Appl. Phys. 37(23), 3296–3303 (2004).
[Crossref]

McConnell, G.

McGhee, E. J.

D. M. Grant, W. Zhang, E. J. McGhee, T. D. Bunney, C. B. Talbot, S. Kumar, I. Munro, C. Dunsby, M. A. Neil, M. Katan, and P. M. French, “Multiplexed FRET to image multiple signaling events in live cells,” Biophys. J. 95(10), L69–L71 (2008).
[Crossref] [PubMed]

McGinty, J.

C. Dunsby, P. M. P. Lanigan, J. McGinty, D. S. Elson, J. Requejo-Isidro, I. Munro, N. Galletly, F. McCann, B. Treanor, B. Önfelt, D. M. Davis, M A A. Neil, and P. M. W. French, “An electronically tunable ultrafast laser source applied to fluorescence imaging and fluorescence lifetime imaging microscopy,” J. Phys. D Appl. Phys. 37(23), 3296–3303 (2004).
[Crossref]

Mitkovski, M.

R. A. Neher, M. Mitkovski, F. Kirchhoff, E. Neher, F. J. Theis, and A. Zeug, “Blind source separation techniques for the decomposition of multiply labeled fluorescence images,” Biophys. J. 96(9), 3791–3800 (2009).
[Crossref] [PubMed]

Moggridge, G. D.

Munro, I.

D. M. Grant, W. Zhang, E. J. McGhee, T. D. Bunney, C. B. Talbot, S. Kumar, I. Munro, C. Dunsby, M. A. Neil, M. Katan, and P. M. French, “Multiplexed FRET to image multiple signaling events in live cells,” Biophys. J. 95(10), L69–L71 (2008).
[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. Önfelt, D. M. Davis, M A A. Neil, and P. M. W. French, “An electronically tunable ultrafast laser source applied to fluorescence imaging and fluorescence lifetime imaging microscopy,” J. Phys. D Appl. Phys. 37(23), 3296–3303 (2004).
[Crossref]

Nagy, P.

D. S. Lidke, P. Nagy, B. G. Barisas, R. Heintzmann, J. N. Post, K. A. Lidke, A. H. Clayton, D. J. Arndt-Jovin, and T. M. Jovin, “Imaging molecular interactions in cells by dynamic and static fluorescence anisotropy (rFLIM and emFRET),” Biochem. Soc. Trans. 31(5), 1020–1027 (2003).
[Crossref] [PubMed]

Neher, E.

R. A. Neher, M. Mitkovski, F. Kirchhoff, E. Neher, F. J. Theis, and A. Zeug, “Blind source separation techniques for the decomposition of multiply labeled fluorescence images,” Biophys. J. 96(9), 3791–3800 (2009).
[Crossref] [PubMed]

Neher, R. A.

R. A. Neher, M. Mitkovski, F. Kirchhoff, E. Neher, F. J. Theis, and A. Zeug, “Blind source separation techniques for the decomposition of multiply labeled fluorescence images,” Biophys. J. 96(9), 3791–3800 (2009).
[Crossref] [PubMed]

Neil, M A A.

C. Dunsby, P. M. P. Lanigan, J. McGinty, D. S. Elson, J. Requejo-Isidro, I. Munro, N. Galletly, F. McCann, B. Treanor, B. Önfelt, D. M. Davis, M A A. Neil, and P. M. W. French, “An electronically tunable ultrafast laser source applied to fluorescence imaging and fluorescence lifetime imaging microscopy,” J. Phys. D Appl. Phys. 37(23), 3296–3303 (2004).
[Crossref]

Neil, M. A.

D. M. Grant, W. Zhang, E. J. McGhee, T. D. Bunney, C. B. Talbot, S. Kumar, I. Munro, C. Dunsby, M. A. Neil, M. Katan, and P. M. French, “Multiplexed FRET to image multiple signaling events in live cells,” Biophys. J. 95(10), L69–L71 (2008).
[Crossref] [PubMed]

H. B. Manning, G. T. Kennedy, D. M. Owen, D. M. Grant, A. I. Magee, M. A. Neil, Y. Itoh, C. Dunsby, and P. M. French, “A compact, multidimensional spectrofluorometer exploiting supercontinuum generation,” J. Biophoton. 1(6), 494 (2008).
[Crossref]

Ng, T.

D. R. Matthews, L. M. Carlin, E. Ofo, P. R. Barber, B. Vojnovic, M. Irving, T. Ng, and S. M. Ameer-Beg, “Time-lapse FRET microscopy using fluorescence anisotropy,” J. Microsc. 237(1), 51–62 (2010).
[Crossref] [PubMed]

Nollen, E. A. A.

T. J. van Ham, A. Esposito, J. R. Kumita, S. T. D. Hsu, G. S. Kaminski Schierle, C. F. Kaminski, C. M. Dobson, E. A. A. Nollen, and C. W. Bertoncini, “Towards multiparametric fluorescent imaging of amyloid formation: studies of a YFP model of alpha-synuclein aggregation,” J. Mol. Biol. 395(3), 627–642 (2010).
[Crossref]

Norris, T. B.

Ofo, E.

D. R. Matthews, L. M. Carlin, E. Ofo, P. R. Barber, B. Vojnovic, M. Irving, T. Ng, and S. M. Ameer-Beg, “Time-lapse FRET microscopy using fluorescence anisotropy,” J. Microsc. 237(1), 51–62 (2010).
[Crossref] [PubMed]

Oggier, T.

A. Esposito, H. C. Gerritsen, T. Oggier, F. Lustenberger, and F. S. Wouters, “Innovating lifetime microscopy: a compact and simple tool for life sciences, screening, and diagnostics,” J. Biomed. Opt. 11(3), 034016 (2006).
[Crossref]

Önfelt, B.

C. Dunsby, P. M. P. Lanigan, J. McGinty, D. S. Elson, J. Requejo-Isidro, I. Munro, N. Galletly, F. McCann, B. Treanor, B. Önfelt, D. M. Davis, M A A. Neil, and P. M. W. French, “An electronically tunable ultrafast laser source applied to fluorescence imaging and fluorescence lifetime imaging microscopy,” J. Phys. D Appl. Phys. 37(23), 3296–3303 (2004).
[Crossref]

Owen, D. M.

H. B. Manning, G. T. Kennedy, D. M. Owen, D. M. Grant, A. I. Magee, M. A. Neil, Y. Itoh, C. Dunsby, and P. M. French, “A compact, multidimensional spectrofluorometer exploiting supercontinuum generation,” J. Biophoton. 1(6), 494 (2008).
[Crossref]

Palero, J. A.

Piston, D. W.

M. A. Rizzo and D. W. Piston, “High-contrast imaging of fluorescent protein FRET by fluorescence polarization microscopy,” Biophys. J. 88(2), L14–L16 (2005).
[Crossref]

Post, J. N.

D. S. Lidke, P. Nagy, B. G. Barisas, R. Heintzmann, J. N. Post, K. A. Lidke, A. H. Clayton, D. J. Arndt-Jovin, and T. M. Jovin, “Imaging molecular interactions in cells by dynamic and static fluorescence anisotropy (rFLIM and emFRET),” Biochem. Soc. Trans. 31(5), 1020–1027 (2003).
[Crossref] [PubMed]

Potier, M.-C.

Rajendra, E.

A. D. Jeyasekharan, N. Ayoub, R. Mahen, J. Ries, A. Esposito, E. Rajendra, H. Hattori, R. P. Kulkarni, and A. R. Venkitaraman, “DNA damage regulates the mobility of Brca2 within the nucleoplasm of living cells,” Proc. Natl. Acad. Sci. U.S.A. 107(50), 21937–21942 (2010).
[Crossref] [PubMed]

Requejo-Isidro, J.

C. Dunsby, P. M. P. Lanigan, J. McGinty, D. S. Elson, J. Requejo-Isidro, I. Munro, N. Galletly, F. McCann, B. Treanor, B. Önfelt, D. M. Davis, M A A. Neil, and P. M. W. French, “An electronically tunable ultrafast laser source applied to fluorescence imaging and fluorescence lifetime imaging microscopy,” J. Phys. D Appl. Phys. 37(23), 3296–3303 (2004).
[Crossref]

Rieger, B.

K. A. Lidke, B. Rieger, D. S. Lidke, and T. M. Jovin, “The role of photon statistics in fluorescence anisotropy imaging,” IEEE Trans. Image Process. 14(9), 1237–1245 (2005).
[Crossref] [PubMed]

Ries, J.

A. D. Jeyasekharan, N. Ayoub, R. Mahen, J. Ries, A. Esposito, E. Rajendra, H. Hattori, R. P. Kulkarni, and A. R. Venkitaraman, “DNA damage regulates the mobility of Brca2 within the nucleoplasm of living cells,” Proc. Natl. Acad. Sci. U.S.A. 107(50), 21937–21942 (2010).
[Crossref] [PubMed]

Rizzo, M. A.

M. A. Rizzo and D. W. Piston, “High-contrast imaging of fluorescent protein FRET by fluorescence polarization microscopy,” Biophys. J. 88(2), L14–L16 (2005).
[Crossref]

Rosen, H.

D. A. Bachovchin, S. J. Brown, H. Rosen, and B. F. Cravatt, “Identification of selective inhibitors of uncharacterized enzymes by high-throughput screening with fluorescent activity-based probes,” Nat. Biotechnol. 27(4), 387–394 (2009).
[Crossref] [PubMed]

Schlachter, S.

Schwedler, S.

Sterenborg, H. J. C. M.

Subramaniam, V.

A. H. A. Clayton, Q. S. Hanley, D. J. Arndt-Jovin, V. Subramaniam, and T. M. Jovin, “Dynamic fluorescence anisotropy imaging microscopy in the frequency domain (rFLIM),” Biophys. J. 83(3), 1631–1649 (2002).
[Crossref] [PubMed]

Swartling, J.

J. H. Frank, A. D. Elder, J. Swartling, A. R. Venkitaraman, A. D. Jeyasekharan, and C. F. Kaminski, “A white light confocal microscope for spectrally resolved multidimensional imaging,” J. Microsc. 227(3), 203–215 (2007).
[Crossref] [PubMed]

Talbot, C. B.

D. M. Grant, W. Zhang, E. J. McGhee, T. D. Bunney, C. B. Talbot, S. Kumar, I. Munro, C. Dunsby, M. A. Neil, M. Katan, and P. M. French, “Multiplexed FRET to image multiple signaling events in live cells,” Biophys. J. 95(10), L69–L71 (2008).
[Crossref] [PubMed]

Theis, F. J.

R. A. Neher, M. Mitkovski, F. Kirchhoff, E. Neher, F. J. Theis, and A. Zeug, “Blind source separation techniques for the decomposition of multiply labeled fluorescence images,” Biophys. J. 96(9), 3791–3800 (2009).
[Crossref] [PubMed]

Tiffert, T.

A. Esposito, T. Tiffert, J. M. Mauritz, S. Schlachter, L. H. Bannister, C. F. Kaminski, and V. L. Lew, “FRET imaging of hemoglobin concentration in Plasmodium falciparum-infected red cells,” PLoS ONE 3(11), e3780 (2008).
[Crossref] [PubMed]

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. Önfelt, D. M. Davis, M A A. Neil, and P. M. W. French, “An electronically tunable ultrafast laser source applied to fluorescence imaging and fluorescence lifetime imaging microscopy,” J. Phys. D Appl. Phys. 37(23), 3296–3303 (2004).
[Crossref]

Valeur, B.

B. Valeur and G. Weber, “Resolution of the fluorescence excitation spectrum of indole into the 1La and 1Lb excitation bands,” Photochem. Photobiol. 25(5), 441–444 (1977).
[Crossref] [PubMed]

van Bergen En Henegouwen, P. M.

van den Heuvel, D. J.

van der Ploeg-van den Heuvel, A.

van Geest, L. K.

van Ham, T. J.

T. J. van Ham, A. Esposito, J. R. Kumita, S. T. D. Hsu, G. S. Kaminski Schierle, C. F. Kaminski, C. M. Dobson, E. A. A. Nollen, and C. W. Bertoncini, “Towards multiparametric fluorescent imaging of amyloid formation: studies of a YFP model of alpha-synuclein aggregation,” J. Mol. Biol. 395(3), 627–642 (2010).
[Crossref]

van Munster, E. B.

G. J. Kremers, E. B. van Munster, J. Goedhart, and T. W. Gadella., “Quantitative lifetime unmixing of multiexponentially decaying fluorophores using single-frequency fluorescence lifetime imaging microscopy,” Biophys. J. 95(1), 378–389 (2008).
[Crossref] [PubMed]

van Sark, W. G. J. H.

Venkitaraman, A. R.

A. D. Jeyasekharan, N. Ayoub, R. Mahen, J. Ries, A. Esposito, E. Rajendra, H. Hattori, R. P. Kulkarni, and A. R. Venkitaraman, “DNA damage regulates the mobility of Brca2 within the nucleoplasm of living cells,” Proc. Natl. Acad. Sci. U.S.A. 107(50), 21937–21942 (2010).
[Crossref] [PubMed]

J. H. Frank, A. D. Elder, J. Swartling, A. R. Venkitaraman, A. D. Jeyasekharan, and C. F. Kaminski, “A white light confocal microscope for spectrally resolved multidimensional imaging,” J. Microsc. 227(3), 203–215 (2007).
[Crossref] [PubMed]

Vojnovic, B.

D. R. Matthews, L. M. Carlin, E. Ofo, P. R. Barber, B. Vojnovic, M. Irving, T. Ng, and S. M. Ameer-Beg, “Time-lapse FRET microscopy using fluorescence anisotropy,” J. Microsc. 237(1), 51–62 (2010).
[Crossref] [PubMed]

Weber, G.

B. Valeur and G. Weber, “Resolution of the fluorescence excitation spectrum of indole into the 1La and 1Lb excitation bands,” Photochem. Photobiol. 25(5), 441–444 (1977).
[Crossref] [PubMed]

Wouters, F. S.

A. Esposito, M. Gralle, M. A. C. Dani, D. Lange, and F. S. Wouters, “pHlameleons: a family of FRET-based protein sensors for quantitative pH imaging,” Biochemistry 47(49), 13115–13126 (2008).
[Crossref] [PubMed]

A. Esposito, H. C. Gerritsen, T. Oggier, F. Lustenberger, and F. S. Wouters, “Innovating lifetime microscopy: a compact and simple tool for life sciences, screening, and diagnostics,” J. Biomed. Opt. 11(3), 034016 (2006).
[Crossref]

Ye, J. Y.

Zamai, M.

M. A. Digman, V. R. Caiolfa, M. Zamai, and E. Gratton, “The phasor approach to fluorescence lifetime imaging analysis,” Biophys. J. 94(2), L14–L16 (2008).
[Crossref]

Zeug, A.

R. A. Neher, M. Mitkovski, F. Kirchhoff, E. Neher, F. J. Theis, and A. Zeug, “Blind source separation techniques for the decomposition of multiply labeled fluorescence images,” Biophys. J. 96(9), 3791–3800 (2009).
[Crossref] [PubMed]

Zhang, W.

D. M. Grant, W. Zhang, E. J. McGhee, T. D. Bunney, C. B. Talbot, S. Kumar, I. Munro, C. Dunsby, M. A. Neil, M. Katan, and P. M. French, “Multiplexed FRET to image multiple signaling events in live cells,” Biophys. J. 95(10), L69–L71 (2008).
[Crossref] [PubMed]

Appl. Opt. (1)

Appl. Spectrosc. (2)

Biochem. Soc. Trans. (1)

D. S. Lidke, P. Nagy, B. G. Barisas, R. Heintzmann, J. N. Post, K. A. Lidke, A. H. Clayton, D. J. Arndt-Jovin, and T. M. Jovin, “Imaging molecular interactions in cells by dynamic and static fluorescence anisotropy (rFLIM and emFRET),” Biochem. Soc. Trans. 31(5), 1020–1027 (2003).
[Crossref] [PubMed]

Biochemistry (1)

A. Esposito, M. Gralle, M. A. C. Dani, D. Lange, and F. S. Wouters, “pHlameleons: a family of FRET-based protein sensors for quantitative pH imaging,” Biochemistry 47(49), 13115–13126 (2008).
[Crossref] [PubMed]

Biophys. J. (6)

G. J. Kremers, E. B. van Munster, J. Goedhart, and T. W. Gadella., “Quantitative lifetime unmixing of multiexponentially decaying fluorophores using single-frequency fluorescence lifetime imaging microscopy,” Biophys. J. 95(1), 378–389 (2008).
[Crossref] [PubMed]

M. A. Rizzo and D. W. Piston, “High-contrast imaging of fluorescent protein FRET by fluorescence polarization microscopy,” Biophys. J. 88(2), L14–L16 (2005).
[Crossref]

A. H. A. Clayton, Q. S. Hanley, D. J. Arndt-Jovin, V. Subramaniam, and T. M. Jovin, “Dynamic fluorescence anisotropy imaging microscopy in the frequency domain (rFLIM),” Biophys. J. 83(3), 1631–1649 (2002).
[Crossref] [PubMed]

R. A. Neher, M. Mitkovski, F. Kirchhoff, E. Neher, F. J. Theis, and A. Zeug, “Blind source separation techniques for the decomposition of multiply labeled fluorescence images,” Biophys. J. 96(9), 3791–3800 (2009).
[Crossref] [PubMed]

D. M. Grant, W. Zhang, E. J. McGhee, T. D. Bunney, C. B. Talbot, S. Kumar, I. Munro, C. Dunsby, M. A. Neil, M. Katan, and P. M. French, “Multiplexed FRET to image multiple signaling events in live cells,” Biophys. J. 95(10), L69–L71 (2008).
[Crossref] [PubMed]

M. A. Digman, V. R. Caiolfa, M. Zamai, and E. Gratton, “The phasor approach to fluorescence lifetime imaging analysis,” Biophys. J. 94(2), L14–L16 (2008).
[Crossref]

ChemPhysChem (1)

F. T. S. Chan, C. F. Kaminski, and G. S. Kaminski Schierle, “HomoFRET fluorescence anisotropy imaging as a tool to study molecular self-assembly in live cells,” ChemPhysChem (2011), doi:.
[Crossref] [PubMed]

IEEE Trans. Image Process. (1)

K. A. Lidke, B. Rieger, D. S. Lidke, and T. M. Jovin, “The role of photon statistics in fluorescence anisotropy imaging,” IEEE Trans. Image Process. 14(9), 1237–1245 (2005).
[Crossref] [PubMed]

J. Biomed. Opt. (1)

A. Esposito, H. C. Gerritsen, T. Oggier, F. Lustenberger, and F. S. Wouters, “Innovating lifetime microscopy: a compact and simple tool for life sciences, screening, and diagnostics,” J. Biomed. Opt. 11(3), 034016 (2006).
[Crossref]

J. Biophoton. (1)

H. B. Manning, G. T. Kennedy, D. M. Owen, D. M. Grant, A. I. Magee, M. A. Neil, Y. Itoh, C. Dunsby, and P. M. French, “A compact, multidimensional spectrofluorometer exploiting supercontinuum generation,” J. Biophoton. 1(6), 494 (2008).
[Crossref]

J. Microsc. (3)

J. H. Frank, A. D. Elder, J. Swartling, A. R. Venkitaraman, A. D. Jeyasekharan, and C. F. Kaminski, “A white light confocal microscope for spectrally resolved multidimensional imaging,” J. Microsc. 227(3), 203–215 (2007).
[Crossref] [PubMed]

D. R. Matthews, L. M. Carlin, E. Ofo, P. R. Barber, B. Vojnovic, M. Irving, T. Ng, and S. M. Ameer-Beg, “Time-lapse FRET microscopy using fluorescence anisotropy,” J. Microsc. 237(1), 51–62 (2010).
[Crossref] [PubMed]

Q. S. Hanley and A. H. Clayton, “AB-plot assisted determination of fluorophore mixtures in a fluorescence lifetime microscope using spectra or quenchers,” J. Microsc. 218(1), 62–67 (2005).
[Crossref] [PubMed]

J. Mol. Biol. (1)

T. J. van Ham, A. Esposito, J. R. Kumita, S. T. D. Hsu, G. S. Kaminski Schierle, C. F. Kaminski, C. M. Dobson, E. A. A. Nollen, and C. W. Bertoncini, “Towards multiparametric fluorescent imaging of amyloid formation: studies of a YFP model of alpha-synuclein aggregation,” J. Mol. Biol. 395(3), 627–642 (2010).
[Crossref]

J. Phys. D Appl. Phys. (1)

C. Dunsby, P. M. P. Lanigan, J. McGinty, D. S. Elson, J. Requejo-Isidro, I. Munro, N. Galletly, F. McCann, B. Treanor, B. Önfelt, D. M. Davis, M A A. Neil, and P. M. W. French, “An electronically tunable ultrafast laser source applied to fluorescence imaging and fluorescence lifetime imaging microscopy,” J. Phys. D Appl. Phys. 37(23), 3296–3303 (2004).
[Crossref]

Nat. Biotechnol. (1)

D. A. Bachovchin, S. J. Brown, H. Rosen, and B. F. Cravatt, “Identification of selective inhibitors of uncharacterized enzymes by high-throughput screening with fluorescent activity-based probes,” Nat. Biotechnol. 27(4), 387–394 (2009).
[Crossref] [PubMed]

Opt. Express (6)

Photochem. Photobiol. (1)

B. Valeur and G. Weber, “Resolution of the fluorescence excitation spectrum of indole into the 1La and 1Lb excitation bands,” Photochem. Photobiol. 25(5), 441–444 (1977).
[Crossref] [PubMed]

PLoS ONE (1)

A. Esposito, T. Tiffert, J. M. Mauritz, S. Schlachter, L. H. Bannister, C. F. Kaminski, and V. L. Lew, “FRET imaging of hemoglobin concentration in Plasmodium falciparum-infected red cells,” PLoS ONE 3(11), e3780 (2008).
[Crossref] [PubMed]

Proc. Natl. Acad. Sci. U.S.A. (1)

A. D. Jeyasekharan, N. Ayoub, R. Mahen, J. Ries, A. Esposito, E. Rajendra, H. Hattori, R. P. Kulkarni, and A. R. Venkitaraman, “DNA damage regulates the mobility of Brca2 within the nucleoplasm of living cells,” Proc. Natl. Acad. Sci. U.S.A. 107(50), 21937–21942 (2010).
[Crossref] [PubMed]

Other (2)

L. Pancheri and D. Stoppa, “A SPAD-based Pixel Linear Array for High-Speed Time-Gated Fluorescence Lifetime Imaging,” 2009 Proc. of Esscirc, 429–432 (2009).

H. C. Gerritsen, A. V. Agronskaia, A. N. Bader, and A. Esposito, “Time Domain FLIM: theory, Instrumentation and data analysis,” in FRET & FLIM Imaging Techniques, T. W. Gadella, ed. (Elsevier, Amsterdam, The Netherlands, 2009).

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 (5)

Fig. 1
Fig. 1 (A) Schematic of principal components in spectropolarimeter; R: half-wave plate retarder; M1-2: silver coated alignment mirrors; L1: collimating lens; O1-2: alignment spots; I1-2: alignment apertures; W: Wollaston prism; P: equilateral dispersive prism; L2: camera lens sc: supercontinuum source and acousto-optical tuneable filter; m1-2: dielectric alignment mirrors; d: dichroic mirror or 80/20 beam splitter; xy: x-y scanners; s: scanning lens; o: objective; t: tube lens; p: confocal pinhole. (b) Typical laser comb used for the calibration of the system with the half-wave plate rotated at 22.5 degrees. (c) Typical spectrum of a fluorescent plastic slide detected on the sensor with the half-plate oriented at 0 degrees.

Download Full Size | PPT Slide | PDF

Fig. 5
Fig. 5 Spectropolarimetry imaging of a FRET pair. (A) Color and anisotropy images of a cell expressing a CFP-17AA-YFP fusion construct (top two rows) are shown when either the donor or the acceptor fluorophores are excited at 440 nm or 490 nm, respectively. When the sample is excited at 490 nm, a 530-550 nm band pass filter was placed before the detector. Also, a cell expressing separated (non-linked) CFP and YFP is shown in the bottom row. Spatially averaged anisotropy spectra (B, top) of these images are shown in red (440 nm excitation) and dark yellow (490 nm excitation). The reference anisotropy spectrum is shown is cyan (no-FRET sample). The ratio between the anisotropy spectra of the samples exhibiting 25% FRET efficiency and the no-FRET sample (B, bottom) clearly show a depolarization caused by FRET present only in the acceptor emission.

Download Full Size | PPT Slide | PDF

Fig. 2
Fig. 2 Spectral and polarization resolved imaging of Convallaria majalis stained with Safranin and Fast Green fluorophores. Panel A shows the average intensity emitted over the entire spectral range upon excitation at 485 nm. B. True color representation of the sample. Panel C shows the anisotropy of the sample overlaid on the intensity map. The information shown in panels B and C is consolidated in panel D. Here, spectral information is contained in the RGB coloring and intensity represents the anisotropy. The bi-dimensional look-up-tables for panel C and D are shown at the bottom. Panels E and F show the spatially averaged emission and anisotropy spectra of the sample.

Download Full Size | PPT Slide | PDF

Fig. 3
Fig. 3 Fluorograms permit the segmentation of HDIM data sets. The fluorescence anisotropy is plotted versus the intensities measured in the red (>570 nm) and green (495-570 nm) spectral regions (A). This fluorogram aided the segmentation of different structures by separating different “clouds” of pixels by gating intensities and anisotropy values (G1-5). The segmented images are shown in panel B; the white arrows show different structures (G4-5) that are segmented based on differences in their anisotropy.

Download Full Size | PPT Slide | PDF

Fig. 4
Fig. 4 Excitation and emission resolved anisotropy spectra. Two regions of interest (yellow and red) of the intensity image are selected (A) and emission spectra are shown as a function of excitation wavelength (B). C summarizes the entire spectral information acquired by the imaging spectropolarimeter in combination with the tuneable supercontinuum source.

Download Full Size | PPT Slide | PDF

Metrics