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Design and application of a confocal microscope for spectrally resolved anisotropy imagingAlessandro Esposito, Arjen N. Bader, Simon C. Schlachter, Dave J. van den Heuvel, Gabriele S. Kaminski Schierle, Ashok R. Venkitaraman, Clemens F. Kaminski, and Hans C. Gerritsen »View Author Affiliations
Alessandro Esposito,1,2,*
Arjen N. Bader,3
Simon C. Schlachter,2
Dave J. van den Heuvel,3
Gabriele S. Kaminski Schierle,2
Ashok R. Venkitaraman,1
Clemens F. Kaminski,2,4
and Hans C. Gerritsen3
1The Medical Research Council Cancer Cell Unit, Hutchison/MRC Research Centre, Hills Road, Cambridge CB2 0XZ, UK 2Department of Chemical Engineering and Biotechnology, University of Cambridge, Pembroke St, Cambridge, CB2 1RA, UK 3Debye Institute, Utrecht University, PO Box 80.000, NL 3508 TA, Utrecht, The Netherlands 4SAOT School of Advanced Optical Technologies, Friedrich Alexander University of Erlangen-Nuremberg, D-91058 Erlangen, Germany *Corresponding author: aesposito@quantitative-microscopy.org |
Optics Express, Vol. 19, Issue 3, pp. 2546-2555 (2011)
http://dx.doi.org/10.1364/OE.19.002546
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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 OSA
OCIS Codes
(170.3880) Medical optics and biotechnology : Medical and biological imaging
(180.1790) Microscopy : Confocal microscopy
(110.4234) Imaging systems : Multispectral and hyperspectral imaging
ToC Category:
Microscopy
History
Original Manuscript: November 16, 2010
Revised Manuscript: January 17, 2011
Manuscript Accepted: January 23, 2011
Published: January 26, 2011
Virtual Issues
Vol. 6, Iss. 2 Virtual Journal for Biomedical Optics
Citation
Alessandro Esposito, Arjen N. Bader, Simon C. Schlachter, Dave J. van den Heuvel, Gabriele S. Kaminski Schierle, Ashok R. Venkitaraman, Clemens F. Kaminski, and Hans C. Gerritsen, "Design and application of a confocal microscope for spectrally resolved anisotropy imaging," Opt. Express 19, 2546-2555 (2011)
http://www.opticsinfobase.org/oe/abstract.cfm?URI=oe-19-3-2546
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References
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- 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]
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- 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]
- 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]
- 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]
- 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. 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]
- 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]
- 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]
- 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]
- 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]
- 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]
- 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]
- 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]
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- 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]
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- 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]
- 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]
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Appl. Opt.
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Biochemistry
- 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.
- 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]
- 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]
- 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]
- 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]
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ChemPhysChem
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IEEE Trans. Image Process.
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J. Biomed. Opt.
- 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.
- 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.
- 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]
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J. Mol. Biol.
- 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.
- 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.
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Opt. Express
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Photochem. Photobiol.
- 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
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Proc. Natl. Acad. Sci. U.S.A.
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Other
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2011, Chan, ChemPhysChem
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- 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]
- 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]
- 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]
- 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]
- 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]
- 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]
- 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]
- 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]
- 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]
- 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]
- 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]
- 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]
- 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]
- 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]
- 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]
- 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]
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