Abstract

In vivo cellular scale fluorescence lifetime imaging of the mouse retina has the potential to be a sensitive marker of retinal cell health. In this study, we demonstrate fluorescence lifetime imaging of extrinsic fluorophores using adaptive optics fluorescence lifetime imaging ophthalmoscopy (AOFLIO). We recorded AOFLIO images of inner retinal cells labeled with enhanced green fluorescent protein (EGFP) and capillaries labeled with fluorescein. We demonstrate that AOFLIO can be used to differentiate spectrally overlapping fluorophores in the retina. With further refinements, AOFLIO could be used to assess retinal health in early stages of degeneration by utilizing lifetime-based sensors or even fluorophores native to the retina.

© 2017 Optical Society of America

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    [Crossref] [PubMed]
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    [Crossref] [PubMed]
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    [Crossref] [PubMed]
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    [Crossref] [PubMed]
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2016 (5)

J. Tam, J. Liu, A. Dubra, and R. Fariss, “In Vivo Imaging of the Human Retinal Pigment Epithelial Mosaic Using Adaptive Optics Enhanced Indocyanine Green Ophthalmoscopy,” Invest. Ophthalmol. Vis. Sci. 57(10), 4376–4384 (2016).
[Crossref] [PubMed]

R. Sharma, D. R. Williams, G. Palczewska, K. Palczewski, and J. J. Hunter, “Two-Photon Autofluorescence Imaging Reveals Cellular Structures Throughout the Retina of the Living Primate Eye,” Invest. Ophthalmol. Vis. Sci. 57(2), 632–646 (2016).
[Crossref] [PubMed]

R. Sharma, C. Schwarz, D. R. Williams, G. Palczewska, K. Palczewski, and J. J. Hunter, “In Vivo Two-Photon Fluorescence Kinetics of Primate Rods and Cones,” Invest. Ophthalmol. Vis. Sci. 57(2), 647–657 (2016).
[Crossref] [PubMed]

C. Dysli, S. Wolf, K. Hatz, and M. S. Zinkernagel, “Fluorescence Lifetime Imaging in Stargardt Disease: Potential Marker for Disease Progression,” Invest. Ophthalmol. Vis. Sci. 57(3), 832–841 (2016).
[Crossref] [PubMed]

A. J. Walsh, J. T. Sharick, M. C. Skala, and H. T. Beier, “Temporal binning of time-correlated single photon counting data improves exponential decay fits and imaging speed,” Biomed. Opt. Express 7(4), 1385–1399 (2016).
[Crossref] [PubMed]

2015 (3)

D. J. Wahl, Y. Jian, S. Bonora, R. J. Zawadzki, and M. V. Sarunic, “Wavefront sensorless adaptive optics fluorescence biomicroscope for in vivo retinal imaging in mice,” Biomed. Opt. Express 7(1), 1–12 (2015).
[Crossref] [PubMed]

M. Klemm, D. Schweitzer, S. Peters, L. Sauer, M. Hammer, and J. Haueisen, “FLIMX: A Software Package to Determine and Analyze the Fluorescence Lifetime in Time-Resolved Fluorescence Data from the Human Eye,” PLoS One 10(7), e0131640 (2015).
[Crossref] [PubMed]

D. Schweitzer, L. Deutsch, M. Klemm, S. Jentsch, M. Hammer, S. Peters, J. Haueisen, U. A. Müller, and J. Dawczynski, “Fluorescence lifetime imaging ophthalmoscopy in type 2 diabetic patients who have no signs of diabetic retinopathy,” J. Biomed. Opt. 20(6), 61106 (2015).
[Crossref] [PubMed]

2014 (5)

T. E. Kornfield and E. A. Newman, “Regulation of Blood Flow in the Retinal Trilaminar Vascular Network,” J. Neurosci. 34(34), 11504–11513 (2014).
[Crossref] [PubMed]

G. Palczewska, Z. Dong, M. Golczak, J. J. Hunter, D. R. Williams, N. S. Alexander, and K. Palczewski, “Noninvasive two-photon microscopy imaging of mouse retina and retinal pigment epithelium through the pupil of the eye,” Nat. Med. 20(7), 785–789 (2014).
[Crossref] [PubMed]

L. Yin, B. Masella, D. Dalkara, J. Zhang, J. G. Flannery, D. V. Schaffer, D. R. Williams, and W. H. Merigan, “Imaging Light Responses of Foveal Ganglion Cells in the Living Macaque Eye,” J. Neurosci. 34(19), 6596–6605 (2014).
[Crossref] [PubMed]

C. Dysli, M. Dysli, V. Enzmann, S. Wolf, and M. S. Zinkernagel, “Fluorescence Lifetime Imaging of the Ocular Fundus in Mice,” Invest. Ophthalmol. Vis. Sci. 55(11), 7206–7215 (2014).
[Crossref] [PubMed]

Q. Yang, J. Zhang, K. Nozato, K. Saito, D. R. Williams, A. Roorda, and E. A. Rossi, “Closed-loop optical stabilization and digital image registration in adaptive optics scanning light ophthalmoscopy,” Biomed. Opt. Express 5(9), 3174–3191 (2014).
[Crossref] [PubMed]

2013 (5)

2012 (1)

2011 (4)

Y. Geng, L. A. Schery, R. Sharma, A. Dubra, K. Ahmad, R. T. Libby, and D. R. Williams, “Optical properties of the mouse eye,” Biomed. Opt. Express 2(4), 717–738 (2011).
[Crossref] [PubMed]

R. W. K. Leung, S.-C. A. Yeh, and Q. Fang, “Effects of incomplete decay in fluorescence lifetime estimation,” Biomed. Opt. Express 2(9), 2517–2531 (2011).
[Crossref] [PubMed]

F. Delori, J. P. Greenberg, R. L. Woods, J. Fischer, T. Duncker, J. Sparrow, and R. T. Smith, “Quantitative measurements of autofluorescence with the scanning laser ophthalmoscope,” Invest. Ophthalmol. Vis. Sci. 52(13), 9379–9390 (2011).
[Crossref] [PubMed]

M. Tantama, Y. P. Hung, and G. Yellen, “Imaging intracellular pH in live cells with a genetically encoded red fluorescent protein sensor,” J. Am. Chem. Soc. 133(26), 10034–10037 (2011).
[Crossref] [PubMed]

2010 (2)

2009 (1)

J. I. W. Morgan, A. Dubra, R. Wolfe, W. H. Merigan, and D. R. Williams, “In Vivo Autofluorescence Imaging of the Human and Macaque Retinal Pigment Epithelial Cell Mosaic,” Invest. Ophthalmol. Vis. Sci. 50(3), 1350–1359 (2009).
[Crossref] [PubMed]

2007 (2)

D. Schweitzer, S. Schenke, M. Hammer, F. Schweitzer, S. Jentsch, E. Birckner, W. Becker, and A. Bergmann, “Towards metabolic mapping of the human retina,” Microsc. Res. Tech. 70(5), 410–419 (2007).
[Crossref] [PubMed]

N. Boens, W. Qin, N. Basarić, J. Hofkens, M. Ameloot, J. Pouget, J.-P. Lefèvre, B. Valeur, E. Gratton, M. vandeVen, N. D. Silva, Y. Engelborghs, K. Willaert, A. Sillen, G. Rumbles, D. Phillips, A. J. Visser, A. van Hoek, J. R. Lakowicz, H. Malak, I. Gryczynski, A. G. Szabo, D. T. Krajcarski, N. Tamai, and A. Miura, “Fluorescence Lifetime Standards for Time and Frequency Domain Fluorescence Spectroscopy,” Anal. Chem. 79(5), 2137–2149 (2007).
[Crossref] [PubMed]

2005 (2)

E. Spiess, F. Bestvater, A. Heckel-Pompey, K. Toth, M. Hacker, G. Stobrawa, T. Feurer, C. Wotzlaw, U. Berchner-Pfannschmidt, T. Porwol, and H. Acker, “Two-photon excitation and emission spectra of the green fluorescent protein variants ECFP, EGFP and EYFP,” J. Microsc. 217(3), 200–204 (2005).
[Crossref] [PubMed]

C. Chen, E. Tsina, M. C. Cornwall, R. K. Crouch, S. Vijayaraghavan, and Y. Koutalos, “Reduction of all-Trans Retinal to All-Trans Retinol in the Outer Segments of Frog and Mouse Rod Photoreceptors,” Biophys. J. 88(3), 2278–2287 (2005).
[Crossref] [PubMed]

2004 (1)

D. Schweitzer, M. Hammer, F. Schweitzer, R. Anders, T. Doebbecke, S. Schenke, E. R. Gaillard, and E. R. Gaillard, “In vivo measurement of time-resolved autofluorescence at the human fundus,” J. Biomed. Opt. 9(6), 1214–1222 (2004).
[Crossref] [PubMed]

2003 (2)

J. R. Lakowicz, J. Malicka, S. D’Auria, and I. Gryczynski, “Release of the self-quenching of fluorescence near silver metallic surfaces,” Anal. Biochem. 320(1), 13–20 (2003).
[Crossref] [PubMed]

S. T. Hess, E. D. Sheets, A. Wagenknecht-Wiesner, and A. A. Heikal, “Quantitative Analysis of the Fluorescence Properties of Intrinsically Fluorescent Proteins in Living Cells,” Biophys. J. 85(4), 2566–2580 (2003).
[Crossref] [PubMed]

2002 (1)

D. Magde, R. Wong, and P. G. Seybold, “Fluorescence Quantum Yields and Their Relation to Lifetimes of Rhodamine 6G and Fluorescein in Nine Solvents: Improved Absolute Standards for Quantum Yields,” Photochem. Photobiol. 75(4), 327–334 (2002).
[Crossref] [PubMed]

2000 (2)

G. Feng, R. H. Mellor, M. Bernstein, C. Keller-Peck, Q. T. Nguyen, M. Wallace, J. M. Nerbonne, J. W. Lichtman, and J. R. Sanes, “Imaging neuronal subsets in transgenic mice expressing multiple spectral variants of GFP,” Neuron 28(1), 41–51 (2000).
[Crossref] [PubMed]

J. Dillon, L. Zheng, J. C. Merriam, and E. R. Gaillard, “Transmission spectra of light to the mammalian retina,” Photochem. Photobiol. 71(2), 225–229 (2000).
[Crossref] [PubMed]

1999 (1)

R. Pepperkok, A. Squire, S. Geley, and P. I. H. Bastiaens, “Simultaneous detection of multiple green fluorescent proteins in live cells by fluorescence lifetime imaging microscopy,” Curr. Biol. 9(5), 269–272 (1999).
[Crossref] [PubMed]

1997 (1)

T. French, P. T. C. So, D. J. Weaver, T. Coelho-Sampaio, E. Gratton, E. W. Voss, and J. Carrero, “Two-photon fluorescence lifetime imaging microscopy of macrophage-mediated antigen processing,” J. Microsc. 185(3), 339–353 (1997).
[Crossref] [PubMed]

1984 (1)

C. J. Barnstable and U. C. Dräger, “Thy-1 antigen: a ganglion cell specific marker in rodent retina,” Neuroscience 11(4), 847–855 (1984).
[Crossref] [PubMed]

Achilefu, S.

M. Y. Berezin and S. Achilefu, “Fluorescence Lifetime Measurements and Biological Imaging,” Chem. Rev. 110(5), 2641–2684 (2010).
[Crossref] [PubMed]

Acker, H.

E. Spiess, F. Bestvater, A. Heckel-Pompey, K. Toth, M. Hacker, G. Stobrawa, T. Feurer, C. Wotzlaw, U. Berchner-Pfannschmidt, T. Porwol, and H. Acker, “Two-photon excitation and emission spectra of the green fluorescent protein variants ECFP, EGFP and EYFP,” J. Microsc. 217(3), 200–204 (2005).
[Crossref] [PubMed]

Ahmad, K.

Alexander, N. S.

G. Palczewska, Z. Dong, M. Golczak, J. J. Hunter, D. R. Williams, N. S. Alexander, and K. Palczewski, “Noninvasive two-photon microscopy imaging of mouse retina and retinal pigment epithelium through the pupil of the eye,” Nat. Med. 20(7), 785–789 (2014).
[Crossref] [PubMed]

Ameloot, M.

N. Boens, W. Qin, N. Basarić, J. Hofkens, M. Ameloot, J. Pouget, J.-P. Lefèvre, B. Valeur, E. Gratton, M. vandeVen, N. D. Silva, Y. Engelborghs, K. Willaert, A. Sillen, G. Rumbles, D. Phillips, A. J. Visser, A. van Hoek, J. R. Lakowicz, H. Malak, I. Gryczynski, A. G. Szabo, D. T. Krajcarski, N. Tamai, and A. Miura, “Fluorescence Lifetime Standards for Time and Frequency Domain Fluorescence Spectroscopy,” Anal. Chem. 79(5), 2137–2149 (2007).
[Crossref] [PubMed]

Anders, R.

D. Schweitzer, M. Hammer, F. Schweitzer, R. Anders, T. Doebbecke, S. Schenke, E. R. Gaillard, and E. R. Gaillard, “In vivo measurement of time-resolved autofluorescence at the human fundus,” J. Biomed. Opt. 9(6), 1214–1222 (2004).
[Crossref] [PubMed]

Balaiya, S.

L. R. Ferguson, J. M. Dominguez, S. Balaiya, S. Grover, and K. V. Chalam, “Retinal Thickness Normative Data in Wild-Type Mice Using Customized Miniature SD-OCT,” PLoS One 8(6), e67265 (2013).
[Crossref] [PubMed]

Barnstable, C. J.

C. J. Barnstable and U. C. Dräger, “Thy-1 antigen: a ganglion cell specific marker in rodent retina,” Neuroscience 11(4), 847–855 (1984).
[Crossref] [PubMed]

Basaric, N.

N. Boens, W. Qin, N. Basarić, J. Hofkens, M. Ameloot, J. Pouget, J.-P. Lefèvre, B. Valeur, E. Gratton, M. vandeVen, N. D. Silva, Y. Engelborghs, K. Willaert, A. Sillen, G. Rumbles, D. Phillips, A. J. Visser, A. van Hoek, J. R. Lakowicz, H. Malak, I. Gryczynski, A. G. Szabo, D. T. Krajcarski, N. Tamai, and A. Miura, “Fluorescence Lifetime Standards for Time and Frequency Domain Fluorescence Spectroscopy,” Anal. Chem. 79(5), 2137–2149 (2007).
[Crossref] [PubMed]

Bastiaens, P. I. H.

R. Pepperkok, A. Squire, S. Geley, and P. I. H. Bastiaens, “Simultaneous detection of multiple green fluorescent proteins in live cells by fluorescence lifetime imaging microscopy,” Curr. Biol. 9(5), 269–272 (1999).
[Crossref] [PubMed]

Becker, W.

D. Schweitzer, S. Schenke, M. Hammer, F. Schweitzer, S. Jentsch, E. Birckner, W. Becker, and A. Bergmann, “Towards metabolic mapping of the human retina,” Microsc. Res. Tech. 70(5), 410–419 (2007).
[Crossref] [PubMed]

Beier, H. T.

Berchner-Pfannschmidt, U.

E. Spiess, F. Bestvater, A. Heckel-Pompey, K. Toth, M. Hacker, G. Stobrawa, T. Feurer, C. Wotzlaw, U. Berchner-Pfannschmidt, T. Porwol, and H. Acker, “Two-photon excitation and emission spectra of the green fluorescent protein variants ECFP, EGFP and EYFP,” J. Microsc. 217(3), 200–204 (2005).
[Crossref] [PubMed]

Berezin, M. Y.

M. Y. Berezin and S. Achilefu, “Fluorescence Lifetime Measurements and Biological Imaging,” Chem. Rev. 110(5), 2641–2684 (2010).
[Crossref] [PubMed]

Bergmann, A.

D. Schweitzer, S. Schenke, M. Hammer, F. Schweitzer, S. Jentsch, E. Birckner, W. Becker, and A. Bergmann, “Towards metabolic mapping of the human retina,” Microsc. Res. Tech. 70(5), 410–419 (2007).
[Crossref] [PubMed]

Bernstein, M.

G. Feng, R. H. Mellor, M. Bernstein, C. Keller-Peck, Q. T. Nguyen, M. Wallace, J. M. Nerbonne, J. W. Lichtman, and J. R. Sanes, “Imaging neuronal subsets in transgenic mice expressing multiple spectral variants of GFP,” Neuron 28(1), 41–51 (2000).
[Crossref] [PubMed]

Bestvater, F.

E. Spiess, F. Bestvater, A. Heckel-Pompey, K. Toth, M. Hacker, G. Stobrawa, T. Feurer, C. Wotzlaw, U. Berchner-Pfannschmidt, T. Porwol, and H. Acker, “Two-photon excitation and emission spectra of the green fluorescent protein variants ECFP, EGFP and EYFP,” J. Microsc. 217(3), 200–204 (2005).
[Crossref] [PubMed]

Birckner, E.

D. Schweitzer, S. Schenke, M. Hammer, F. Schweitzer, S. Jentsch, E. Birckner, W. Becker, and A. Bergmann, “Towards metabolic mapping of the human retina,” Microsc. Res. Tech. 70(5), 410–419 (2007).
[Crossref] [PubMed]

Boens, N.

N. Boens, W. Qin, N. Basarić, J. Hofkens, M. Ameloot, J. Pouget, J.-P. Lefèvre, B. Valeur, E. Gratton, M. vandeVen, N. D. Silva, Y. Engelborghs, K. Willaert, A. Sillen, G. Rumbles, D. Phillips, A. J. Visser, A. van Hoek, J. R. Lakowicz, H. Malak, I. Gryczynski, A. G. Szabo, D. T. Krajcarski, N. Tamai, and A. Miura, “Fluorescence Lifetime Standards for Time and Frequency Domain Fluorescence Spectroscopy,” Anal. Chem. 79(5), 2137–2149 (2007).
[Crossref] [PubMed]

Bonora, S.

Callaway, E. M.

L. Yin, Y. Geng, F. Osakada, R. Sharma, A. H. Cetin, E. M. Callaway, D. R. Williams, and W. H. Merigan, “Imaging light responses of retinal ganglion cells in the living mouse eye,” J. Neurophysiol. 109(9), 2415–2421 (2013).
[Crossref] [PubMed]

Carrero, J.

T. French, P. T. C. So, D. J. Weaver, T. Coelho-Sampaio, E. Gratton, E. W. Voss, and J. Carrero, “Two-photon fluorescence lifetime imaging microscopy of macrophage-mediated antigen processing,” J. Microsc. 185(3), 339–353 (1997).
[Crossref] [PubMed]

Carroll, J.

Cetin, A. H.

L. Yin, Y. Geng, F. Osakada, R. Sharma, A. H. Cetin, E. M. Callaway, D. R. Williams, and W. H. Merigan, “Imaging light responses of retinal ganglion cells in the living mouse eye,” J. Neurophysiol. 109(9), 2415–2421 (2013).
[Crossref] [PubMed]

Chalam, K. V.

L. R. Ferguson, J. M. Dominguez, S. Balaiya, S. Grover, and K. V. Chalam, “Retinal Thickness Normative Data in Wild-Type Mice Using Customized Miniature SD-OCT,” PLoS One 8(6), e67265 (2013).
[Crossref] [PubMed]

Chen, C.

C. Chen, E. Tsina, M. C. Cornwall, R. K. Crouch, S. Vijayaraghavan, and Y. Koutalos, “Reduction of all-Trans Retinal to All-Trans Retinol in the Outer Segments of Frog and Mouse Rod Photoreceptors,” Biophys. J. 88(3), 2278–2287 (2005).
[Crossref] [PubMed]

Chui, T. Y.

Chung, M. M.

Coelho-Sampaio, T.

T. French, P. T. C. So, D. J. Weaver, T. Coelho-Sampaio, E. Gratton, E. W. Voss, and J. Carrero, “Two-photon fluorescence lifetime imaging microscopy of macrophage-mediated antigen processing,” J. Microsc. 185(3), 339–353 (1997).
[Crossref] [PubMed]

Cornwall, M. C.

C. Chen, E. Tsina, M. C. Cornwall, R. K. Crouch, S. Vijayaraghavan, and Y. Koutalos, “Reduction of all-Trans Retinal to All-Trans Retinol in the Outer Segments of Frog and Mouse Rod Photoreceptors,” Biophys. J. 88(3), 2278–2287 (2005).
[Crossref] [PubMed]

Crouch, R. K.

C. Chen, E. Tsina, M. C. Cornwall, R. K. Crouch, S. Vijayaraghavan, and Y. Koutalos, “Reduction of all-Trans Retinal to All-Trans Retinol in the Outer Segments of Frog and Mouse Rod Photoreceptors,” Biophys. J. 88(3), 2278–2287 (2005).
[Crossref] [PubMed]

D’Auria, S.

J. R. Lakowicz, J. Malicka, S. D’Auria, and I. Gryczynski, “Release of the self-quenching of fluorescence near silver metallic surfaces,” Anal. Biochem. 320(1), 13–20 (2003).
[Crossref] [PubMed]

Dalkara, D.

L. Yin, B. Masella, D. Dalkara, J. Zhang, J. G. Flannery, D. V. Schaffer, D. R. Williams, and W. H. Merigan, “Imaging Light Responses of Foveal Ganglion Cells in the Living Macaque Eye,” J. Neurosci. 34(19), 6596–6605 (2014).
[Crossref] [PubMed]

Dawczynski, J.

D. Schweitzer, L. Deutsch, M. Klemm, S. Jentsch, M. Hammer, S. Peters, J. Haueisen, U. A. Müller, and J. Dawczynski, “Fluorescence lifetime imaging ophthalmoscopy in type 2 diabetic patients who have no signs of diabetic retinopathy,” J. Biomed. Opt. 20(6), 61106 (2015).
[Crossref] [PubMed]

Delori, F.

F. Delori, J. P. Greenberg, R. L. Woods, J. Fischer, T. Duncker, J. Sparrow, and R. T. Smith, “Quantitative measurements of autofluorescence with the scanning laser ophthalmoscope,” Invest. Ophthalmol. Vis. Sci. 52(13), 9379–9390 (2011).
[Crossref] [PubMed]

Deutsch, L.

D. Schweitzer, L. Deutsch, M. Klemm, S. Jentsch, M. Hammer, S. Peters, J. Haueisen, U. A. Müller, and J. Dawczynski, “Fluorescence lifetime imaging ophthalmoscopy in type 2 diabetic patients who have no signs of diabetic retinopathy,” J. Biomed. Opt. 20(6), 61106 (2015).
[Crossref] [PubMed]

Dillon, J.

J. Dillon, L. Zheng, J. C. Merriam, and E. R. Gaillard, “Transmission spectra of light to the mammalian retina,” Photochem. Photobiol. 71(2), 225–229 (2000).
[Crossref] [PubMed]

Doebbecke, T.

D. Schweitzer, M. Hammer, F. Schweitzer, R. Anders, T. Doebbecke, S. Schenke, E. R. Gaillard, and E. R. Gaillard, “In vivo measurement of time-resolved autofluorescence at the human fundus,” J. Biomed. Opt. 9(6), 1214–1222 (2004).
[Crossref] [PubMed]

Dominguez, J. M.

L. R. Ferguson, J. M. Dominguez, S. Balaiya, S. Grover, and K. V. Chalam, “Retinal Thickness Normative Data in Wild-Type Mice Using Customized Miniature SD-OCT,” PLoS One 8(6), e67265 (2013).
[Crossref] [PubMed]

Dong, Z.

G. Palczewska, Z. Dong, M. Golczak, J. J. Hunter, D. R. Williams, N. S. Alexander, and K. Palczewski, “Noninvasive two-photon microscopy imaging of mouse retina and retinal pigment epithelium through the pupil of the eye,” Nat. Med. 20(7), 785–789 (2014).
[Crossref] [PubMed]

Dräger, U. C.

C. J. Barnstable and U. C. Dräger, “Thy-1 antigen: a ganglion cell specific marker in rodent retina,” Neuroscience 11(4), 847–855 (1984).
[Crossref] [PubMed]

Dubow, M.

Dubra, A.

J. Tam, J. Liu, A. Dubra, and R. Fariss, “In Vivo Imaging of the Human Retinal Pigment Epithelial Mosaic Using Adaptive Optics Enhanced Indocyanine Green Ophthalmoscopy,” Invest. Ophthalmol. Vis. Sci. 57(10), 4376–4384 (2016).
[Crossref] [PubMed]

A. Pinhas, M. Dubow, N. Shah, T. Y. Chui, D. Scoles, Y. N. Sulai, R. Weitz, J. B. Walsh, J. Carroll, A. Dubra, and R. B. Rosen, “In vivo imaging of human retinal microvasculature using adaptive optics scanning light ophthalmoscope fluorescein angiography,” Biomed. Opt. Express 4(8), 1305–1317 (2013).
[Crossref] [PubMed]

E. A. Rossi, P. Rangel-Fonseca, K. Parkins, W. Fischer, L. R. Latchney, M. A. Folwell, D. R. Williams, A. Dubra, and M. M. Chung, “In vivo imaging of retinal pigment epithelium cells in age related macular degeneration,” Biomed. Opt. Express 4(11), 2527–2539 (2013).
[Crossref] [PubMed]

Y. Geng, A. Dubra, L. Yin, W. H. Merigan, R. Sharma, R. T. Libby, and D. R. Williams, “Adaptive optics retinal imaging in the living mouse eye,” Biomed. Opt. Express 3(4), 715–734 (2012).
[Crossref] [PubMed]

Y. Geng, L. A. Schery, R. Sharma, A. Dubra, K. Ahmad, R. T. Libby, and D. R. Williams, “Optical properties of the mouse eye,” Biomed. Opt. Express 2(4), 717–738 (2011).
[Crossref] [PubMed]

J. J. Hunter, B. Masella, A. Dubra, R. Sharma, L. Yin, W. H. Merigan, G. Palczewska, K. Palczewski, and D. R. Williams, “Images of photoreceptors in living primate eyes using adaptive optics two-photon ophthalmoscopy,” Biomed. Opt. Express 2(1), 139–148 (2010).
[Crossref] [PubMed]

J. I. W. Morgan, A. Dubra, R. Wolfe, W. H. Merigan, and D. R. Williams, “In Vivo Autofluorescence Imaging of the Human and Macaque Retinal Pigment Epithelial Cell Mosaic,” Invest. Ophthalmol. Vis. Sci. 50(3), 1350–1359 (2009).
[Crossref] [PubMed]

Duncker, T.

F. Delori, J. P. Greenberg, R. L. Woods, J. Fischer, T. Duncker, J. Sparrow, and R. T. Smith, “Quantitative measurements of autofluorescence with the scanning laser ophthalmoscope,” Invest. Ophthalmol. Vis. Sci. 52(13), 9379–9390 (2011).
[Crossref] [PubMed]

Dysli, C.

C. Dysli, S. Wolf, K. Hatz, and M. S. Zinkernagel, “Fluorescence Lifetime Imaging in Stargardt Disease: Potential Marker for Disease Progression,” Invest. Ophthalmol. Vis. Sci. 57(3), 832–841 (2016).
[Crossref] [PubMed]

C. Dysli, M. Dysli, V. Enzmann, S. Wolf, and M. S. Zinkernagel, “Fluorescence Lifetime Imaging of the Ocular Fundus in Mice,” Invest. Ophthalmol. Vis. Sci. 55(11), 7206–7215 (2014).
[Crossref] [PubMed]

Dysli, M.

C. Dysli, M. Dysli, V. Enzmann, S. Wolf, and M. S. Zinkernagel, “Fluorescence Lifetime Imaging of the Ocular Fundus in Mice,” Invest. Ophthalmol. Vis. Sci. 55(11), 7206–7215 (2014).
[Crossref] [PubMed]

Engelborghs, Y.

N. Boens, W. Qin, N. Basarić, J. Hofkens, M. Ameloot, J. Pouget, J.-P. Lefèvre, B. Valeur, E. Gratton, M. vandeVen, N. D. Silva, Y. Engelborghs, K. Willaert, A. Sillen, G. Rumbles, D. Phillips, A. J. Visser, A. van Hoek, J. R. Lakowicz, H. Malak, I. Gryczynski, A. G. Szabo, D. T. Krajcarski, N. Tamai, and A. Miura, “Fluorescence Lifetime Standards for Time and Frequency Domain Fluorescence Spectroscopy,” Anal. Chem. 79(5), 2137–2149 (2007).
[Crossref] [PubMed]

Enzmann, V.

C. Dysli, M. Dysli, V. Enzmann, S. Wolf, and M. S. Zinkernagel, “Fluorescence Lifetime Imaging of the Ocular Fundus in Mice,” Invest. Ophthalmol. Vis. Sci. 55(11), 7206–7215 (2014).
[Crossref] [PubMed]

Fang, Q.

Fariss, R.

J. Tam, J. Liu, A. Dubra, and R. Fariss, “In Vivo Imaging of the Human Retinal Pigment Epithelial Mosaic Using Adaptive Optics Enhanced Indocyanine Green Ophthalmoscopy,” Invest. Ophthalmol. Vis. Sci. 57(10), 4376–4384 (2016).
[Crossref] [PubMed]

Feng, G.

G. Feng, R. H. Mellor, M. Bernstein, C. Keller-Peck, Q. T. Nguyen, M. Wallace, J. M. Nerbonne, J. W. Lichtman, and J. R. Sanes, “Imaging neuronal subsets in transgenic mice expressing multiple spectral variants of GFP,” Neuron 28(1), 41–51 (2000).
[Crossref] [PubMed]

Ferguson, L. R.

L. R. Ferguson, J. M. Dominguez, S. Balaiya, S. Grover, and K. V. Chalam, “Retinal Thickness Normative Data in Wild-Type Mice Using Customized Miniature SD-OCT,” PLoS One 8(6), e67265 (2013).
[Crossref] [PubMed]

Feurer, T.

E. Spiess, F. Bestvater, A. Heckel-Pompey, K. Toth, M. Hacker, G. Stobrawa, T. Feurer, C. Wotzlaw, U. Berchner-Pfannschmidt, T. Porwol, and H. Acker, “Two-photon excitation and emission spectra of the green fluorescent protein variants ECFP, EGFP and EYFP,” J. Microsc. 217(3), 200–204 (2005).
[Crossref] [PubMed]

Fischer, J.

F. Delori, J. P. Greenberg, R. L. Woods, J. Fischer, T. Duncker, J. Sparrow, and R. T. Smith, “Quantitative measurements of autofluorescence with the scanning laser ophthalmoscope,” Invest. Ophthalmol. Vis. Sci. 52(13), 9379–9390 (2011).
[Crossref] [PubMed]

Fischer, W.

Flannery, J. G.

L. Yin, B. Masella, D. Dalkara, J. Zhang, J. G. Flannery, D. V. Schaffer, D. R. Williams, and W. H. Merigan, “Imaging Light Responses of Foveal Ganglion Cells in the Living Macaque Eye,” J. Neurosci. 34(19), 6596–6605 (2014).
[Crossref] [PubMed]

Folwell, M. A.

French, T.

T. French, P. T. C. So, D. J. Weaver, T. Coelho-Sampaio, E. Gratton, E. W. Voss, and J. Carrero, “Two-photon fluorescence lifetime imaging microscopy of macrophage-mediated antigen processing,” J. Microsc. 185(3), 339–353 (1997).
[Crossref] [PubMed]

Gaillard, E. R.

D. Schweitzer, M. Hammer, F. Schweitzer, R. Anders, T. Doebbecke, S. Schenke, E. R. Gaillard, and E. R. Gaillard, “In vivo measurement of time-resolved autofluorescence at the human fundus,” J. Biomed. Opt. 9(6), 1214–1222 (2004).
[Crossref] [PubMed]

D. Schweitzer, M. Hammer, F. Schweitzer, R. Anders, T. Doebbecke, S. Schenke, E. R. Gaillard, and E. R. Gaillard, “In vivo measurement of time-resolved autofluorescence at the human fundus,” J. Biomed. Opt. 9(6), 1214–1222 (2004).
[Crossref] [PubMed]

J. Dillon, L. Zheng, J. C. Merriam, and E. R. Gaillard, “Transmission spectra of light to the mammalian retina,” Photochem. Photobiol. 71(2), 225–229 (2000).
[Crossref] [PubMed]

Geley, S.

R. Pepperkok, A. Squire, S. Geley, and P. I. H. Bastiaens, “Simultaneous detection of multiple green fluorescent proteins in live cells by fluorescence lifetime imaging microscopy,” Curr. Biol. 9(5), 269–272 (1999).
[Crossref] [PubMed]

Geng, Y.

Golczak, M.

G. Palczewska, Z. Dong, M. Golczak, J. J. Hunter, D. R. Williams, N. S. Alexander, and K. Palczewski, “Noninvasive two-photon microscopy imaging of mouse retina and retinal pigment epithelium through the pupil of the eye,” Nat. Med. 20(7), 785–789 (2014).
[Crossref] [PubMed]

Gratton, E.

N. Boens, W. Qin, N. Basarić, J. Hofkens, M. Ameloot, J. Pouget, J.-P. Lefèvre, B. Valeur, E. Gratton, M. vandeVen, N. D. Silva, Y. Engelborghs, K. Willaert, A. Sillen, G. Rumbles, D. Phillips, A. J. Visser, A. van Hoek, J. R. Lakowicz, H. Malak, I. Gryczynski, A. G. Szabo, D. T. Krajcarski, N. Tamai, and A. Miura, “Fluorescence Lifetime Standards for Time and Frequency Domain Fluorescence Spectroscopy,” Anal. Chem. 79(5), 2137–2149 (2007).
[Crossref] [PubMed]

T. French, P. T. C. So, D. J. Weaver, T. Coelho-Sampaio, E. Gratton, E. W. Voss, and J. Carrero, “Two-photon fluorescence lifetime imaging microscopy of macrophage-mediated antigen processing,” J. Microsc. 185(3), 339–353 (1997).
[Crossref] [PubMed]

Greenberg, J. P.

F. Delori, J. P. Greenberg, R. L. Woods, J. Fischer, T. Duncker, J. Sparrow, and R. T. Smith, “Quantitative measurements of autofluorescence with the scanning laser ophthalmoscope,” Invest. Ophthalmol. Vis. Sci. 52(13), 9379–9390 (2011).
[Crossref] [PubMed]

Grover, S.

L. R. Ferguson, J. M. Dominguez, S. Balaiya, S. Grover, and K. V. Chalam, “Retinal Thickness Normative Data in Wild-Type Mice Using Customized Miniature SD-OCT,” PLoS One 8(6), e67265 (2013).
[Crossref] [PubMed]

Gryczynski, I.

N. Boens, W. Qin, N. Basarić, J. Hofkens, M. Ameloot, J. Pouget, J.-P. Lefèvre, B. Valeur, E. Gratton, M. vandeVen, N. D. Silva, Y. Engelborghs, K. Willaert, A. Sillen, G. Rumbles, D. Phillips, A. J. Visser, A. van Hoek, J. R. Lakowicz, H. Malak, I. Gryczynski, A. G. Szabo, D. T. Krajcarski, N. Tamai, and A. Miura, “Fluorescence Lifetime Standards for Time and Frequency Domain Fluorescence Spectroscopy,” Anal. Chem. 79(5), 2137–2149 (2007).
[Crossref] [PubMed]

J. R. Lakowicz, J. Malicka, S. D’Auria, and I. Gryczynski, “Release of the self-quenching of fluorescence near silver metallic surfaces,” Anal. Biochem. 320(1), 13–20 (2003).
[Crossref] [PubMed]

Hacker, M.

E. Spiess, F. Bestvater, A. Heckel-Pompey, K. Toth, M. Hacker, G. Stobrawa, T. Feurer, C. Wotzlaw, U. Berchner-Pfannschmidt, T. Porwol, and H. Acker, “Two-photon excitation and emission spectra of the green fluorescent protein variants ECFP, EGFP and EYFP,” J. Microsc. 217(3), 200–204 (2005).
[Crossref] [PubMed]

Hammer, M.

D. Schweitzer, L. Deutsch, M. Klemm, S. Jentsch, M. Hammer, S. Peters, J. Haueisen, U. A. Müller, and J. Dawczynski, “Fluorescence lifetime imaging ophthalmoscopy in type 2 diabetic patients who have no signs of diabetic retinopathy,” J. Biomed. Opt. 20(6), 61106 (2015).
[Crossref] [PubMed]

M. Klemm, D. Schweitzer, S. Peters, L. Sauer, M. Hammer, and J. Haueisen, “FLIMX: A Software Package to Determine and Analyze the Fluorescence Lifetime in Time-Resolved Fluorescence Data from the Human Eye,” PLoS One 10(7), e0131640 (2015).
[Crossref] [PubMed]

D. Schweitzer, S. Schenke, M. Hammer, F. Schweitzer, S. Jentsch, E. Birckner, W. Becker, and A. Bergmann, “Towards metabolic mapping of the human retina,” Microsc. Res. Tech. 70(5), 410–419 (2007).
[Crossref] [PubMed]

D. Schweitzer, M. Hammer, F. Schweitzer, R. Anders, T. Doebbecke, S. Schenke, E. R. Gaillard, and E. R. Gaillard, “In vivo measurement of time-resolved autofluorescence at the human fundus,” J. Biomed. Opt. 9(6), 1214–1222 (2004).
[Crossref] [PubMed]

Hatz, K.

C. Dysli, S. Wolf, K. Hatz, and M. S. Zinkernagel, “Fluorescence Lifetime Imaging in Stargardt Disease: Potential Marker for Disease Progression,” Invest. Ophthalmol. Vis. Sci. 57(3), 832–841 (2016).
[Crossref] [PubMed]

Haueisen, J.

M. Klemm, D. Schweitzer, S. Peters, L. Sauer, M. Hammer, and J. Haueisen, “FLIMX: A Software Package to Determine and Analyze the Fluorescence Lifetime in Time-Resolved Fluorescence Data from the Human Eye,” PLoS One 10(7), e0131640 (2015).
[Crossref] [PubMed]

D. Schweitzer, L. Deutsch, M. Klemm, S. Jentsch, M. Hammer, S. Peters, J. Haueisen, U. A. Müller, and J. Dawczynski, “Fluorescence lifetime imaging ophthalmoscopy in type 2 diabetic patients who have no signs of diabetic retinopathy,” J. Biomed. Opt. 20(6), 61106 (2015).
[Crossref] [PubMed]

Heckel-Pompey, A.

E. Spiess, F. Bestvater, A. Heckel-Pompey, K. Toth, M. Hacker, G. Stobrawa, T. Feurer, C. Wotzlaw, U. Berchner-Pfannschmidt, T. Porwol, and H. Acker, “Two-photon excitation and emission spectra of the green fluorescent protein variants ECFP, EGFP and EYFP,” J. Microsc. 217(3), 200–204 (2005).
[Crossref] [PubMed]

Heikal, A. A.

S. T. Hess, E. D. Sheets, A. Wagenknecht-Wiesner, and A. A. Heikal, “Quantitative Analysis of the Fluorescence Properties of Intrinsically Fluorescent Proteins in Living Cells,” Biophys. J. 85(4), 2566–2580 (2003).
[Crossref] [PubMed]

Hess, S. T.

S. T. Hess, E. D. Sheets, A. Wagenknecht-Wiesner, and A. A. Heikal, “Quantitative Analysis of the Fluorescence Properties of Intrinsically Fluorescent Proteins in Living Cells,” Biophys. J. 85(4), 2566–2580 (2003).
[Crossref] [PubMed]

Hofkens, J.

N. Boens, W. Qin, N. Basarić, J. Hofkens, M. Ameloot, J. Pouget, J.-P. Lefèvre, B. Valeur, E. Gratton, M. vandeVen, N. D. Silva, Y. Engelborghs, K. Willaert, A. Sillen, G. Rumbles, D. Phillips, A. J. Visser, A. van Hoek, J. R. Lakowicz, H. Malak, I. Gryczynski, A. G. Szabo, D. T. Krajcarski, N. Tamai, and A. Miura, “Fluorescence Lifetime Standards for Time and Frequency Domain Fluorescence Spectroscopy,” Anal. Chem. 79(5), 2137–2149 (2007).
[Crossref] [PubMed]

Hung, Y. P.

M. Tantama, Y. P. Hung, and G. Yellen, “Imaging intracellular pH in live cells with a genetically encoded red fluorescent protein sensor,” J. Am. Chem. Soc. 133(26), 10034–10037 (2011).
[Crossref] [PubMed]

Hunter, J. J.

R. Sharma, D. R. Williams, G. Palczewska, K. Palczewski, and J. J. Hunter, “Two-Photon Autofluorescence Imaging Reveals Cellular Structures Throughout the Retina of the Living Primate Eye,” Invest. Ophthalmol. Vis. Sci. 57(2), 632–646 (2016).
[Crossref] [PubMed]

R. Sharma, C. Schwarz, D. R. Williams, G. Palczewska, K. Palczewski, and J. J. Hunter, “In Vivo Two-Photon Fluorescence Kinetics of Primate Rods and Cones,” Invest. Ophthalmol. Vis. Sci. 57(2), 647–657 (2016).
[Crossref] [PubMed]

G. Palczewska, Z. Dong, M. Golczak, J. J. Hunter, D. R. Williams, N. S. Alexander, and K. Palczewski, “Noninvasive two-photon microscopy imaging of mouse retina and retinal pigment epithelium through the pupil of the eye,” Nat. Med. 20(7), 785–789 (2014).
[Crossref] [PubMed]

R. Sharma, L. Yin, Y. Geng, W. H. Merigan, G. Palczewska, K. Palczewski, D. R. Williams, and J. J. Hunter, “In vivo two-photon imaging of the mouse retina,” Biomed. Opt. Express 4(8), 1285–1293 (2013).
[Crossref] [PubMed]

J. J. Hunter, B. Masella, A. Dubra, R. Sharma, L. Yin, W. H. Merigan, G. Palczewska, K. Palczewski, and D. R. Williams, “Images of photoreceptors in living primate eyes using adaptive optics two-photon ophthalmoscopy,” Biomed. Opt. Express 2(1), 139–148 (2010).
[Crossref] [PubMed]

Jentsch, S.

D. Schweitzer, L. Deutsch, M. Klemm, S. Jentsch, M. Hammer, S. Peters, J. Haueisen, U. A. Müller, and J. Dawczynski, “Fluorescence lifetime imaging ophthalmoscopy in type 2 diabetic patients who have no signs of diabetic retinopathy,” J. Biomed. Opt. 20(6), 61106 (2015).
[Crossref] [PubMed]

D. Schweitzer, S. Schenke, M. Hammer, F. Schweitzer, S. Jentsch, E. Birckner, W. Becker, and A. Bergmann, “Towards metabolic mapping of the human retina,” Microsc. Res. Tech. 70(5), 410–419 (2007).
[Crossref] [PubMed]

Jian, Y.

Keller-Peck, C.

G. Feng, R. H. Mellor, M. Bernstein, C. Keller-Peck, Q. T. Nguyen, M. Wallace, J. M. Nerbonne, J. W. Lichtman, and J. R. Sanes, “Imaging neuronal subsets in transgenic mice expressing multiple spectral variants of GFP,” Neuron 28(1), 41–51 (2000).
[Crossref] [PubMed]

Klemm, M.

M. Klemm, D. Schweitzer, S. Peters, L. Sauer, M. Hammer, and J. Haueisen, “FLIMX: A Software Package to Determine and Analyze the Fluorescence Lifetime in Time-Resolved Fluorescence Data from the Human Eye,” PLoS One 10(7), e0131640 (2015).
[Crossref] [PubMed]

D. Schweitzer, L. Deutsch, M. Klemm, S. Jentsch, M. Hammer, S. Peters, J. Haueisen, U. A. Müller, and J. Dawczynski, “Fluorescence lifetime imaging ophthalmoscopy in type 2 diabetic patients who have no signs of diabetic retinopathy,” J. Biomed. Opt. 20(6), 61106 (2015).
[Crossref] [PubMed]

Kornfield, T. E.

T. E. Kornfield and E. A. Newman, “Regulation of Blood Flow in the Retinal Trilaminar Vascular Network,” J. Neurosci. 34(34), 11504–11513 (2014).
[Crossref] [PubMed]

Koutalos, Y.

C. Chen, E. Tsina, M. C. Cornwall, R. K. Crouch, S. Vijayaraghavan, and Y. Koutalos, “Reduction of all-Trans Retinal to All-Trans Retinol in the Outer Segments of Frog and Mouse Rod Photoreceptors,” Biophys. J. 88(3), 2278–2287 (2005).
[Crossref] [PubMed]

Krajcarski, D. T.

N. Boens, W. Qin, N. Basarić, J. Hofkens, M. Ameloot, J. Pouget, J.-P. Lefèvre, B. Valeur, E. Gratton, M. vandeVen, N. D. Silva, Y. Engelborghs, K. Willaert, A. Sillen, G. Rumbles, D. Phillips, A. J. Visser, A. van Hoek, J. R. Lakowicz, H. Malak, I. Gryczynski, A. G. Szabo, D. T. Krajcarski, N. Tamai, and A. Miura, “Fluorescence Lifetime Standards for Time and Frequency Domain Fluorescence Spectroscopy,” Anal. Chem. 79(5), 2137–2149 (2007).
[Crossref] [PubMed]

Lakowicz, J. R.

N. Boens, W. Qin, N. Basarić, J. Hofkens, M. Ameloot, J. Pouget, J.-P. Lefèvre, B. Valeur, E. Gratton, M. vandeVen, N. D. Silva, Y. Engelborghs, K. Willaert, A. Sillen, G. Rumbles, D. Phillips, A. J. Visser, A. van Hoek, J. R. Lakowicz, H. Malak, I. Gryczynski, A. G. Szabo, D. T. Krajcarski, N. Tamai, and A. Miura, “Fluorescence Lifetime Standards for Time and Frequency Domain Fluorescence Spectroscopy,” Anal. Chem. 79(5), 2137–2149 (2007).
[Crossref] [PubMed]

J. R. Lakowicz, J. Malicka, S. D’Auria, and I. Gryczynski, “Release of the self-quenching of fluorescence near silver metallic surfaces,” Anal. Biochem. 320(1), 13–20 (2003).
[Crossref] [PubMed]

Latchney, L. R.

Lefèvre, J.-P.

N. Boens, W. Qin, N. Basarić, J. Hofkens, M. Ameloot, J. Pouget, J.-P. Lefèvre, B. Valeur, E. Gratton, M. vandeVen, N. D. Silva, Y. Engelborghs, K. Willaert, A. Sillen, G. Rumbles, D. Phillips, A. J. Visser, A. van Hoek, J. R. Lakowicz, H. Malak, I. Gryczynski, A. G. Szabo, D. T. Krajcarski, N. Tamai, and A. Miura, “Fluorescence Lifetime Standards for Time and Frequency Domain Fluorescence Spectroscopy,” Anal. Chem. 79(5), 2137–2149 (2007).
[Crossref] [PubMed]

Leung, R. W. K.

Libby, R. T.

Lichtman, J. W.

G. Feng, R. H. Mellor, M. Bernstein, C. Keller-Peck, Q. T. Nguyen, M. Wallace, J. M. Nerbonne, J. W. Lichtman, and J. R. Sanes, “Imaging neuronal subsets in transgenic mice expressing multiple spectral variants of GFP,” Neuron 28(1), 41–51 (2000).
[Crossref] [PubMed]

Liu, J.

J. Tam, J. Liu, A. Dubra, and R. Fariss, “In Vivo Imaging of the Human Retinal Pigment Epithelial Mosaic Using Adaptive Optics Enhanced Indocyanine Green Ophthalmoscopy,” Invest. Ophthalmol. Vis. Sci. 57(10), 4376–4384 (2016).
[Crossref] [PubMed]

Magde, D.

D. Magde, R. Wong, and P. G. Seybold, “Fluorescence Quantum Yields and Their Relation to Lifetimes of Rhodamine 6G and Fluorescein in Nine Solvents: Improved Absolute Standards for Quantum Yields,” Photochem. Photobiol. 75(4), 327–334 (2002).
[Crossref] [PubMed]

Malak, H.

N. Boens, W. Qin, N. Basarić, J. Hofkens, M. Ameloot, J. Pouget, J.-P. Lefèvre, B. Valeur, E. Gratton, M. vandeVen, N. D. Silva, Y. Engelborghs, K. Willaert, A. Sillen, G. Rumbles, D. Phillips, A. J. Visser, A. van Hoek, J. R. Lakowicz, H. Malak, I. Gryczynski, A. G. Szabo, D. T. Krajcarski, N. Tamai, and A. Miura, “Fluorescence Lifetime Standards for Time and Frequency Domain Fluorescence Spectroscopy,” Anal. Chem. 79(5), 2137–2149 (2007).
[Crossref] [PubMed]

Malicka, J.

J. R. Lakowicz, J. Malicka, S. D’Auria, and I. Gryczynski, “Release of the self-quenching of fluorescence near silver metallic surfaces,” Anal. Biochem. 320(1), 13–20 (2003).
[Crossref] [PubMed]

Masella, B.

L. Yin, B. Masella, D. Dalkara, J. Zhang, J. G. Flannery, D. V. Schaffer, D. R. Williams, and W. H. Merigan, “Imaging Light Responses of Foveal Ganglion Cells in the Living Macaque Eye,” J. Neurosci. 34(19), 6596–6605 (2014).
[Crossref] [PubMed]

J. J. Hunter, B. Masella, A. Dubra, R. Sharma, L. Yin, W. H. Merigan, G. Palczewska, K. Palczewski, and D. R. Williams, “Images of photoreceptors in living primate eyes using adaptive optics two-photon ophthalmoscopy,” Biomed. Opt. Express 2(1), 139–148 (2010).
[Crossref] [PubMed]

Mellor, R. H.

G. Feng, R. H. Mellor, M. Bernstein, C. Keller-Peck, Q. T. Nguyen, M. Wallace, J. M. Nerbonne, J. W. Lichtman, and J. R. Sanes, “Imaging neuronal subsets in transgenic mice expressing multiple spectral variants of GFP,” Neuron 28(1), 41–51 (2000).
[Crossref] [PubMed]

Merigan, W. H.

L. Yin, B. Masella, D. Dalkara, J. Zhang, J. G. Flannery, D. V. Schaffer, D. R. Williams, and W. H. Merigan, “Imaging Light Responses of Foveal Ganglion Cells in the Living Macaque Eye,” J. Neurosci. 34(19), 6596–6605 (2014).
[Crossref] [PubMed]

R. Sharma, L. Yin, Y. Geng, W. H. Merigan, G. Palczewska, K. Palczewski, D. R. Williams, and J. J. Hunter, “In vivo two-photon imaging of the mouse retina,” Biomed. Opt. Express 4(8), 1285–1293 (2013).
[Crossref] [PubMed]

L. Yin, Y. Geng, F. Osakada, R. Sharma, A. H. Cetin, E. M. Callaway, D. R. Williams, and W. H. Merigan, “Imaging light responses of retinal ganglion cells in the living mouse eye,” J. Neurophysiol. 109(9), 2415–2421 (2013).
[Crossref] [PubMed]

Y. Geng, A. Dubra, L. Yin, W. H. Merigan, R. Sharma, R. T. Libby, and D. R. Williams, “Adaptive optics retinal imaging in the living mouse eye,” Biomed. Opt. Express 3(4), 715–734 (2012).
[Crossref] [PubMed]

J. J. Hunter, B. Masella, A. Dubra, R. Sharma, L. Yin, W. H. Merigan, G. Palczewska, K. Palczewski, and D. R. Williams, “Images of photoreceptors in living primate eyes using adaptive optics two-photon ophthalmoscopy,” Biomed. Opt. Express 2(1), 139–148 (2010).
[Crossref] [PubMed]

J. I. W. Morgan, A. Dubra, R. Wolfe, W. H. Merigan, and D. R. Williams, “In Vivo Autofluorescence Imaging of the Human and Macaque Retinal Pigment Epithelial Cell Mosaic,” Invest. Ophthalmol. Vis. Sci. 50(3), 1350–1359 (2009).
[Crossref] [PubMed]

Merriam, J. C.

J. Dillon, L. Zheng, J. C. Merriam, and E. R. Gaillard, “Transmission spectra of light to the mammalian retina,” Photochem. Photobiol. 71(2), 225–229 (2000).
[Crossref] [PubMed]

Miura, A.

N. Boens, W. Qin, N. Basarić, J. Hofkens, M. Ameloot, J. Pouget, J.-P. Lefèvre, B. Valeur, E. Gratton, M. vandeVen, N. D. Silva, Y. Engelborghs, K. Willaert, A. Sillen, G. Rumbles, D. Phillips, A. J. Visser, A. van Hoek, J. R. Lakowicz, H. Malak, I. Gryczynski, A. G. Szabo, D. T. Krajcarski, N. Tamai, and A. Miura, “Fluorescence Lifetime Standards for Time and Frequency Domain Fluorescence Spectroscopy,” Anal. Chem. 79(5), 2137–2149 (2007).
[Crossref] [PubMed]

Morgan, J. I. W.

J. I. W. Morgan, A. Dubra, R. Wolfe, W. H. Merigan, and D. R. Williams, “In Vivo Autofluorescence Imaging of the Human and Macaque Retinal Pigment Epithelial Cell Mosaic,” Invest. Ophthalmol. Vis. Sci. 50(3), 1350–1359 (2009).
[Crossref] [PubMed]

Müller, U. A.

D. Schweitzer, L. Deutsch, M. Klemm, S. Jentsch, M. Hammer, S. Peters, J. Haueisen, U. A. Müller, and J. Dawczynski, “Fluorescence lifetime imaging ophthalmoscopy in type 2 diabetic patients who have no signs of diabetic retinopathy,” J. Biomed. Opt. 20(6), 61106 (2015).
[Crossref] [PubMed]

Nerbonne, J. M.

G. Feng, R. H. Mellor, M. Bernstein, C. Keller-Peck, Q. T. Nguyen, M. Wallace, J. M. Nerbonne, J. W. Lichtman, and J. R. Sanes, “Imaging neuronal subsets in transgenic mice expressing multiple spectral variants of GFP,” Neuron 28(1), 41–51 (2000).
[Crossref] [PubMed]

Newman, E. A.

T. E. Kornfield and E. A. Newman, “Regulation of Blood Flow in the Retinal Trilaminar Vascular Network,” J. Neurosci. 34(34), 11504–11513 (2014).
[Crossref] [PubMed]

Nguyen, Q. T.

G. Feng, R. H. Mellor, M. Bernstein, C. Keller-Peck, Q. T. Nguyen, M. Wallace, J. M. Nerbonne, J. W. Lichtman, and J. R. Sanes, “Imaging neuronal subsets in transgenic mice expressing multiple spectral variants of GFP,” Neuron 28(1), 41–51 (2000).
[Crossref] [PubMed]

Nozato, K.

Osakada, F.

L. Yin, Y. Geng, F. Osakada, R. Sharma, A. H. Cetin, E. M. Callaway, D. R. Williams, and W. H. Merigan, “Imaging light responses of retinal ganglion cells in the living mouse eye,” J. Neurophysiol. 109(9), 2415–2421 (2013).
[Crossref] [PubMed]

Palczewska, G.

R. Sharma, C. Schwarz, D. R. Williams, G. Palczewska, K. Palczewski, and J. J. Hunter, “In Vivo Two-Photon Fluorescence Kinetics of Primate Rods and Cones,” Invest. Ophthalmol. Vis. Sci. 57(2), 647–657 (2016).
[Crossref] [PubMed]

R. Sharma, D. R. Williams, G. Palczewska, K. Palczewski, and J. J. Hunter, “Two-Photon Autofluorescence Imaging Reveals Cellular Structures Throughout the Retina of the Living Primate Eye,” Invest. Ophthalmol. Vis. Sci. 57(2), 632–646 (2016).
[Crossref] [PubMed]

G. Palczewska, Z. Dong, M. Golczak, J. J. Hunter, D. R. Williams, N. S. Alexander, and K. Palczewski, “Noninvasive two-photon microscopy imaging of mouse retina and retinal pigment epithelium through the pupil of the eye,” Nat. Med. 20(7), 785–789 (2014).
[Crossref] [PubMed]

R. Sharma, L. Yin, Y. Geng, W. H. Merigan, G. Palczewska, K. Palczewski, D. R. Williams, and J. J. Hunter, “In vivo two-photon imaging of the mouse retina,” Biomed. Opt. Express 4(8), 1285–1293 (2013).
[Crossref] [PubMed]

J. J. Hunter, B. Masella, A. Dubra, R. Sharma, L. Yin, W. H. Merigan, G. Palczewska, K. Palczewski, and D. R. Williams, “Images of photoreceptors in living primate eyes using adaptive optics two-photon ophthalmoscopy,” Biomed. Opt. Express 2(1), 139–148 (2010).
[Crossref] [PubMed]

Palczewski, K.

R. Sharma, D. R. Williams, G. Palczewska, K. Palczewski, and J. J. Hunter, “Two-Photon Autofluorescence Imaging Reveals Cellular Structures Throughout the Retina of the Living Primate Eye,” Invest. Ophthalmol. Vis. Sci. 57(2), 632–646 (2016).
[Crossref] [PubMed]

R. Sharma, C. Schwarz, D. R. Williams, G. Palczewska, K. Palczewski, and J. J. Hunter, “In Vivo Two-Photon Fluorescence Kinetics of Primate Rods and Cones,” Invest. Ophthalmol. Vis. Sci. 57(2), 647–657 (2016).
[Crossref] [PubMed]

G. Palczewska, Z. Dong, M. Golczak, J. J. Hunter, D. R. Williams, N. S. Alexander, and K. Palczewski, “Noninvasive two-photon microscopy imaging of mouse retina and retinal pigment epithelium through the pupil of the eye,” Nat. Med. 20(7), 785–789 (2014).
[Crossref] [PubMed]

R. Sharma, L. Yin, Y. Geng, W. H. Merigan, G. Palczewska, K. Palczewski, D. R. Williams, and J. J. Hunter, “In vivo two-photon imaging of the mouse retina,” Biomed. Opt. Express 4(8), 1285–1293 (2013).
[Crossref] [PubMed]

J. J. Hunter, B. Masella, A. Dubra, R. Sharma, L. Yin, W. H. Merigan, G. Palczewska, K. Palczewski, and D. R. Williams, “Images of photoreceptors in living primate eyes using adaptive optics two-photon ophthalmoscopy,” Biomed. Opt. Express 2(1), 139–148 (2010).
[Crossref] [PubMed]

Parkins, K.

Pepperkok, R.

R. Pepperkok, A. Squire, S. Geley, and P. I. H. Bastiaens, “Simultaneous detection of multiple green fluorescent proteins in live cells by fluorescence lifetime imaging microscopy,” Curr. Biol. 9(5), 269–272 (1999).
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Peters, S.

D. Schweitzer, L. Deutsch, M. Klemm, S. Jentsch, M. Hammer, S. Peters, J. Haueisen, U. A. Müller, and J. Dawczynski, “Fluorescence lifetime imaging ophthalmoscopy in type 2 diabetic patients who have no signs of diabetic retinopathy,” J. Biomed. Opt. 20(6), 61106 (2015).
[Crossref] [PubMed]

M. Klemm, D. Schweitzer, S. Peters, L. Sauer, M. Hammer, and J. Haueisen, “FLIMX: A Software Package to Determine and Analyze the Fluorescence Lifetime in Time-Resolved Fluorescence Data from the Human Eye,” PLoS One 10(7), e0131640 (2015).
[Crossref] [PubMed]

Phillips, D.

N. Boens, W. Qin, N. Basarić, J. Hofkens, M. Ameloot, J. Pouget, J.-P. Lefèvre, B. Valeur, E. Gratton, M. vandeVen, N. D. Silva, Y. Engelborghs, K. Willaert, A. Sillen, G. Rumbles, D. Phillips, A. J. Visser, A. van Hoek, J. R. Lakowicz, H. Malak, I. Gryczynski, A. G. Szabo, D. T. Krajcarski, N. Tamai, and A. Miura, “Fluorescence Lifetime Standards for Time and Frequency Domain Fluorescence Spectroscopy,” Anal. Chem. 79(5), 2137–2149 (2007).
[Crossref] [PubMed]

Pinhas, A.

Porwol, T.

E. Spiess, F. Bestvater, A. Heckel-Pompey, K. Toth, M. Hacker, G. Stobrawa, T. Feurer, C. Wotzlaw, U. Berchner-Pfannschmidt, T. Porwol, and H. Acker, “Two-photon excitation and emission spectra of the green fluorescent protein variants ECFP, EGFP and EYFP,” J. Microsc. 217(3), 200–204 (2005).
[Crossref] [PubMed]

Pouget, J.

N. Boens, W. Qin, N. Basarić, J. Hofkens, M. Ameloot, J. Pouget, J.-P. Lefèvre, B. Valeur, E. Gratton, M. vandeVen, N. D. Silva, Y. Engelborghs, K. Willaert, A. Sillen, G. Rumbles, D. Phillips, A. J. Visser, A. van Hoek, J. R. Lakowicz, H. Malak, I. Gryczynski, A. G. Szabo, D. T. Krajcarski, N. Tamai, and A. Miura, “Fluorescence Lifetime Standards for Time and Frequency Domain Fluorescence Spectroscopy,” Anal. Chem. 79(5), 2137–2149 (2007).
[Crossref] [PubMed]

Qin, W.

N. Boens, W. Qin, N. Basarić, J. Hofkens, M. Ameloot, J. Pouget, J.-P. Lefèvre, B. Valeur, E. Gratton, M. vandeVen, N. D. Silva, Y. Engelborghs, K. Willaert, A. Sillen, G. Rumbles, D. Phillips, A. J. Visser, A. van Hoek, J. R. Lakowicz, H. Malak, I. Gryczynski, A. G. Szabo, D. T. Krajcarski, N. Tamai, and A. Miura, “Fluorescence Lifetime Standards for Time and Frequency Domain Fluorescence Spectroscopy,” Anal. Chem. 79(5), 2137–2149 (2007).
[Crossref] [PubMed]

Rangel-Fonseca, P.

Roorda, A.

Rosen, R. B.

Rossi, E. A.

Rumbles, G.

N. Boens, W. Qin, N. Basarić, J. Hofkens, M. Ameloot, J. Pouget, J.-P. Lefèvre, B. Valeur, E. Gratton, M. vandeVen, N. D. Silva, Y. Engelborghs, K. Willaert, A. Sillen, G. Rumbles, D. Phillips, A. J. Visser, A. van Hoek, J. R. Lakowicz, H. Malak, I. Gryczynski, A. G. Szabo, D. T. Krajcarski, N. Tamai, and A. Miura, “Fluorescence Lifetime Standards for Time and Frequency Domain Fluorescence Spectroscopy,” Anal. Chem. 79(5), 2137–2149 (2007).
[Crossref] [PubMed]

Saito, K.

Sanes, J. R.

G. Feng, R. H. Mellor, M. Bernstein, C. Keller-Peck, Q. T. Nguyen, M. Wallace, J. M. Nerbonne, J. W. Lichtman, and J. R. Sanes, “Imaging neuronal subsets in transgenic mice expressing multiple spectral variants of GFP,” Neuron 28(1), 41–51 (2000).
[Crossref] [PubMed]

Sarunic, M. V.

Sauer, L.

M. Klemm, D. Schweitzer, S. Peters, L. Sauer, M. Hammer, and J. Haueisen, “FLIMX: A Software Package to Determine and Analyze the Fluorescence Lifetime in Time-Resolved Fluorescence Data from the Human Eye,” PLoS One 10(7), e0131640 (2015).
[Crossref] [PubMed]

Schaffer, D. V.

L. Yin, B. Masella, D. Dalkara, J. Zhang, J. G. Flannery, D. V. Schaffer, D. R. Williams, and W. H. Merigan, “Imaging Light Responses of Foveal Ganglion Cells in the Living Macaque Eye,” J. Neurosci. 34(19), 6596–6605 (2014).
[Crossref] [PubMed]

Schenke, S.

D. Schweitzer, S. Schenke, M. Hammer, F. Schweitzer, S. Jentsch, E. Birckner, W. Becker, and A. Bergmann, “Towards metabolic mapping of the human retina,” Microsc. Res. Tech. 70(5), 410–419 (2007).
[Crossref] [PubMed]

D. Schweitzer, M. Hammer, F. Schweitzer, R. Anders, T. Doebbecke, S. Schenke, E. R. Gaillard, and E. R. Gaillard, “In vivo measurement of time-resolved autofluorescence at the human fundus,” J. Biomed. Opt. 9(6), 1214–1222 (2004).
[Crossref] [PubMed]

Schery, L. A.

Schwarz, C.

R. Sharma, C. Schwarz, D. R. Williams, G. Palczewska, K. Palczewski, and J. J. Hunter, “In Vivo Two-Photon Fluorescence Kinetics of Primate Rods and Cones,” Invest. Ophthalmol. Vis. Sci. 57(2), 647–657 (2016).
[Crossref] [PubMed]

Schweitzer, D.

M. Klemm, D. Schweitzer, S. Peters, L. Sauer, M. Hammer, and J. Haueisen, “FLIMX: A Software Package to Determine and Analyze the Fluorescence Lifetime in Time-Resolved Fluorescence Data from the Human Eye,” PLoS One 10(7), e0131640 (2015).
[Crossref] [PubMed]

D. Schweitzer, L. Deutsch, M. Klemm, S. Jentsch, M. Hammer, S. Peters, J. Haueisen, U. A. Müller, and J. Dawczynski, “Fluorescence lifetime imaging ophthalmoscopy in type 2 diabetic patients who have no signs of diabetic retinopathy,” J. Biomed. Opt. 20(6), 61106 (2015).
[Crossref] [PubMed]

D. Schweitzer, S. Schenke, M. Hammer, F. Schweitzer, S. Jentsch, E. Birckner, W. Becker, and A. Bergmann, “Towards metabolic mapping of the human retina,” Microsc. Res. Tech. 70(5), 410–419 (2007).
[Crossref] [PubMed]

D. Schweitzer, M. Hammer, F. Schweitzer, R. Anders, T. Doebbecke, S. Schenke, E. R. Gaillard, and E. R. Gaillard, “In vivo measurement of time-resolved autofluorescence at the human fundus,” J. Biomed. Opt. 9(6), 1214–1222 (2004).
[Crossref] [PubMed]

Schweitzer, F.

D. Schweitzer, S. Schenke, M. Hammer, F. Schweitzer, S. Jentsch, E. Birckner, W. Becker, and A. Bergmann, “Towards metabolic mapping of the human retina,” Microsc. Res. Tech. 70(5), 410–419 (2007).
[Crossref] [PubMed]

D. Schweitzer, M. Hammer, F. Schweitzer, R. Anders, T. Doebbecke, S. Schenke, E. R. Gaillard, and E. R. Gaillard, “In vivo measurement of time-resolved autofluorescence at the human fundus,” J. Biomed. Opt. 9(6), 1214–1222 (2004).
[Crossref] [PubMed]

Scoles, D.

Seybold, P. G.

D. Magde, R. Wong, and P. G. Seybold, “Fluorescence Quantum Yields and Their Relation to Lifetimes of Rhodamine 6G and Fluorescein in Nine Solvents: Improved Absolute Standards for Quantum Yields,” Photochem. Photobiol. 75(4), 327–334 (2002).
[Crossref] [PubMed]

Shah, N.

Sharick, J. T.

Sharma, R.

R. Sharma, C. Schwarz, D. R. Williams, G. Palczewska, K. Palczewski, and J. J. Hunter, “In Vivo Two-Photon Fluorescence Kinetics of Primate Rods and Cones,” Invest. Ophthalmol. Vis. Sci. 57(2), 647–657 (2016).
[Crossref] [PubMed]

R. Sharma, D. R. Williams, G. Palczewska, K. Palczewski, and J. J. Hunter, “Two-Photon Autofluorescence Imaging Reveals Cellular Structures Throughout the Retina of the Living Primate Eye,” Invest. Ophthalmol. Vis. Sci. 57(2), 632–646 (2016).
[Crossref] [PubMed]

L. Yin, Y. Geng, F. Osakada, R. Sharma, A. H. Cetin, E. M. Callaway, D. R. Williams, and W. H. Merigan, “Imaging light responses of retinal ganglion cells in the living mouse eye,” J. Neurophysiol. 109(9), 2415–2421 (2013).
[Crossref] [PubMed]

R. Sharma, L. Yin, Y. Geng, W. H. Merigan, G. Palczewska, K. Palczewski, D. R. Williams, and J. J. Hunter, “In vivo two-photon imaging of the mouse retina,” Biomed. Opt. Express 4(8), 1285–1293 (2013).
[Crossref] [PubMed]

Y. Geng, A. Dubra, L. Yin, W. H. Merigan, R. Sharma, R. T. Libby, and D. R. Williams, “Adaptive optics retinal imaging in the living mouse eye,” Biomed. Opt. Express 3(4), 715–734 (2012).
[Crossref] [PubMed]

Y. Geng, L. A. Schery, R. Sharma, A. Dubra, K. Ahmad, R. T. Libby, and D. R. Williams, “Optical properties of the mouse eye,” Biomed. Opt. Express 2(4), 717–738 (2011).
[Crossref] [PubMed]

J. J. Hunter, B. Masella, A. Dubra, R. Sharma, L. Yin, W. H. Merigan, G. Palczewska, K. Palczewski, and D. R. Williams, “Images of photoreceptors in living primate eyes using adaptive optics two-photon ophthalmoscopy,” Biomed. Opt. Express 2(1), 139–148 (2010).
[Crossref] [PubMed]

Sheets, E. D.

S. T. Hess, E. D. Sheets, A. Wagenknecht-Wiesner, and A. A. Heikal, “Quantitative Analysis of the Fluorescence Properties of Intrinsically Fluorescent Proteins in Living Cells,” Biophys. J. 85(4), 2566–2580 (2003).
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Sillen, A.

N. Boens, W. Qin, N. Basarić, J. Hofkens, M. Ameloot, J. Pouget, J.-P. Lefèvre, B. Valeur, E. Gratton, M. vandeVen, N. D. Silva, Y. Engelborghs, K. Willaert, A. Sillen, G. Rumbles, D. Phillips, A. J. Visser, A. van Hoek, J. R. Lakowicz, H. Malak, I. Gryczynski, A. G. Szabo, D. T. Krajcarski, N. Tamai, and A. Miura, “Fluorescence Lifetime Standards for Time and Frequency Domain Fluorescence Spectroscopy,” Anal. Chem. 79(5), 2137–2149 (2007).
[Crossref] [PubMed]

Silva, N. D.

N. Boens, W. Qin, N. Basarić, J. Hofkens, M. Ameloot, J. Pouget, J.-P. Lefèvre, B. Valeur, E. Gratton, M. vandeVen, N. D. Silva, Y. Engelborghs, K. Willaert, A. Sillen, G. Rumbles, D. Phillips, A. J. Visser, A. van Hoek, J. R. Lakowicz, H. Malak, I. Gryczynski, A. G. Szabo, D. T. Krajcarski, N. Tamai, and A. Miura, “Fluorescence Lifetime Standards for Time and Frequency Domain Fluorescence Spectroscopy,” Anal. Chem. 79(5), 2137–2149 (2007).
[Crossref] [PubMed]

Skala, M. C.

Smith, R. T.

F. Delori, J. P. Greenberg, R. L. Woods, J. Fischer, T. Duncker, J. Sparrow, and R. T. Smith, “Quantitative measurements of autofluorescence with the scanning laser ophthalmoscope,” Invest. Ophthalmol. Vis. Sci. 52(13), 9379–9390 (2011).
[Crossref] [PubMed]

So, P. T. C.

T. French, P. T. C. So, D. J. Weaver, T. Coelho-Sampaio, E. Gratton, E. W. Voss, and J. Carrero, “Two-photon fluorescence lifetime imaging microscopy of macrophage-mediated antigen processing,” J. Microsc. 185(3), 339–353 (1997).
[Crossref] [PubMed]

Sparrow, J.

F. Delori, J. P. Greenberg, R. L. Woods, J. Fischer, T. Duncker, J. Sparrow, and R. T. Smith, “Quantitative measurements of autofluorescence with the scanning laser ophthalmoscope,” Invest. Ophthalmol. Vis. Sci. 52(13), 9379–9390 (2011).
[Crossref] [PubMed]

Spiess, E.

E. Spiess, F. Bestvater, A. Heckel-Pompey, K. Toth, M. Hacker, G. Stobrawa, T. Feurer, C. Wotzlaw, U. Berchner-Pfannschmidt, T. Porwol, and H. Acker, “Two-photon excitation and emission spectra of the green fluorescent protein variants ECFP, EGFP and EYFP,” J. Microsc. 217(3), 200–204 (2005).
[Crossref] [PubMed]

Squire, A.

R. Pepperkok, A. Squire, S. Geley, and P. I. H. Bastiaens, “Simultaneous detection of multiple green fluorescent proteins in live cells by fluorescence lifetime imaging microscopy,” Curr. Biol. 9(5), 269–272 (1999).
[Crossref] [PubMed]

Stobrawa, G.

E. Spiess, F. Bestvater, A. Heckel-Pompey, K. Toth, M. Hacker, G. Stobrawa, T. Feurer, C. Wotzlaw, U. Berchner-Pfannschmidt, T. Porwol, and H. Acker, “Two-photon excitation and emission spectra of the green fluorescent protein variants ECFP, EGFP and EYFP,” J. Microsc. 217(3), 200–204 (2005).
[Crossref] [PubMed]

Sulai, Y. N.

Szabo, A. G.

N. Boens, W. Qin, N. Basarić, J. Hofkens, M. Ameloot, J. Pouget, J.-P. Lefèvre, B. Valeur, E. Gratton, M. vandeVen, N. D. Silva, Y. Engelborghs, K. Willaert, A. Sillen, G. Rumbles, D. Phillips, A. J. Visser, A. van Hoek, J. R. Lakowicz, H. Malak, I. Gryczynski, A. G. Szabo, D. T. Krajcarski, N. Tamai, and A. Miura, “Fluorescence Lifetime Standards for Time and Frequency Domain Fluorescence Spectroscopy,” Anal. Chem. 79(5), 2137–2149 (2007).
[Crossref] [PubMed]

Tam, J.

J. Tam, J. Liu, A. Dubra, and R. Fariss, “In Vivo Imaging of the Human Retinal Pigment Epithelial Mosaic Using Adaptive Optics Enhanced Indocyanine Green Ophthalmoscopy,” Invest. Ophthalmol. Vis. Sci. 57(10), 4376–4384 (2016).
[Crossref] [PubMed]

Tamai, N.

N. Boens, W. Qin, N. Basarić, J. Hofkens, M. Ameloot, J. Pouget, J.-P. Lefèvre, B. Valeur, E. Gratton, M. vandeVen, N. D. Silva, Y. Engelborghs, K. Willaert, A. Sillen, G. Rumbles, D. Phillips, A. J. Visser, A. van Hoek, J. R. Lakowicz, H. Malak, I. Gryczynski, A. G. Szabo, D. T. Krajcarski, N. Tamai, and A. Miura, “Fluorescence Lifetime Standards for Time and Frequency Domain Fluorescence Spectroscopy,” Anal. Chem. 79(5), 2137–2149 (2007).
[Crossref] [PubMed]

Tantama, M.

M. Tantama, Y. P. Hung, and G. Yellen, “Imaging intracellular pH in live cells with a genetically encoded red fluorescent protein sensor,” J. Am. Chem. Soc. 133(26), 10034–10037 (2011).
[Crossref] [PubMed]

Toth, K.

E. Spiess, F. Bestvater, A. Heckel-Pompey, K. Toth, M. Hacker, G. Stobrawa, T. Feurer, C. Wotzlaw, U. Berchner-Pfannschmidt, T. Porwol, and H. Acker, “Two-photon excitation and emission spectra of the green fluorescent protein variants ECFP, EGFP and EYFP,” J. Microsc. 217(3), 200–204 (2005).
[Crossref] [PubMed]

Tsina, E.

C. Chen, E. Tsina, M. C. Cornwall, R. K. Crouch, S. Vijayaraghavan, and Y. Koutalos, “Reduction of all-Trans Retinal to All-Trans Retinol in the Outer Segments of Frog and Mouse Rod Photoreceptors,” Biophys. J. 88(3), 2278–2287 (2005).
[Crossref] [PubMed]

Valeur, B.

N. Boens, W. Qin, N. Basarić, J. Hofkens, M. Ameloot, J. Pouget, J.-P. Lefèvre, B. Valeur, E. Gratton, M. vandeVen, N. D. Silva, Y. Engelborghs, K. Willaert, A. Sillen, G. Rumbles, D. Phillips, A. J. Visser, A. van Hoek, J. R. Lakowicz, H. Malak, I. Gryczynski, A. G. Szabo, D. T. Krajcarski, N. Tamai, and A. Miura, “Fluorescence Lifetime Standards for Time and Frequency Domain Fluorescence Spectroscopy,” Anal. Chem. 79(5), 2137–2149 (2007).
[Crossref] [PubMed]

van Hoek, A.

N. Boens, W. Qin, N. Basarić, J. Hofkens, M. Ameloot, J. Pouget, J.-P. Lefèvre, B. Valeur, E. Gratton, M. vandeVen, N. D. Silva, Y. Engelborghs, K. Willaert, A. Sillen, G. Rumbles, D. Phillips, A. J. Visser, A. van Hoek, J. R. Lakowicz, H. Malak, I. Gryczynski, A. G. Szabo, D. T. Krajcarski, N. Tamai, and A. Miura, “Fluorescence Lifetime Standards for Time and Frequency Domain Fluorescence Spectroscopy,” Anal. Chem. 79(5), 2137–2149 (2007).
[Crossref] [PubMed]

vandeVen, M.

N. Boens, W. Qin, N. Basarić, J. Hofkens, M. Ameloot, J. Pouget, J.-P. Lefèvre, B. Valeur, E. Gratton, M. vandeVen, N. D. Silva, Y. Engelborghs, K. Willaert, A. Sillen, G. Rumbles, D. Phillips, A. J. Visser, A. van Hoek, J. R. Lakowicz, H. Malak, I. Gryczynski, A. G. Szabo, D. T. Krajcarski, N. Tamai, and A. Miura, “Fluorescence Lifetime Standards for Time and Frequency Domain Fluorescence Spectroscopy,” Anal. Chem. 79(5), 2137–2149 (2007).
[Crossref] [PubMed]

Vijayaraghavan, S.

C. Chen, E. Tsina, M. C. Cornwall, R. K. Crouch, S. Vijayaraghavan, and Y. Koutalos, “Reduction of all-Trans Retinal to All-Trans Retinol in the Outer Segments of Frog and Mouse Rod Photoreceptors,” Biophys. J. 88(3), 2278–2287 (2005).
[Crossref] [PubMed]

Visser, A. J.

N. Boens, W. Qin, N. Basarić, J. Hofkens, M. Ameloot, J. Pouget, J.-P. Lefèvre, B. Valeur, E. Gratton, M. vandeVen, N. D. Silva, Y. Engelborghs, K. Willaert, A. Sillen, G. Rumbles, D. Phillips, A. J. Visser, A. van Hoek, J. R. Lakowicz, H. Malak, I. Gryczynski, A. G. Szabo, D. T. Krajcarski, N. Tamai, and A. Miura, “Fluorescence Lifetime Standards for Time and Frequency Domain Fluorescence Spectroscopy,” Anal. Chem. 79(5), 2137–2149 (2007).
[Crossref] [PubMed]

Voss, E. W.

T. French, P. T. C. So, D. J. Weaver, T. Coelho-Sampaio, E. Gratton, E. W. Voss, and J. Carrero, “Two-photon fluorescence lifetime imaging microscopy of macrophage-mediated antigen processing,” J. Microsc. 185(3), 339–353 (1997).
[Crossref] [PubMed]

Wagenknecht-Wiesner, A.

S. T. Hess, E. D. Sheets, A. Wagenknecht-Wiesner, and A. A. Heikal, “Quantitative Analysis of the Fluorescence Properties of Intrinsically Fluorescent Proteins in Living Cells,” Biophys. J. 85(4), 2566–2580 (2003).
[Crossref] [PubMed]

Wahl, D. J.

Wallace, M.

G. Feng, R. H. Mellor, M. Bernstein, C. Keller-Peck, Q. T. Nguyen, M. Wallace, J. M. Nerbonne, J. W. Lichtman, and J. R. Sanes, “Imaging neuronal subsets in transgenic mice expressing multiple spectral variants of GFP,” Neuron 28(1), 41–51 (2000).
[Crossref] [PubMed]

Walsh, A. J.

Walsh, J. B.

Weaver, D. J.

T. French, P. T. C. So, D. J. Weaver, T. Coelho-Sampaio, E. Gratton, E. W. Voss, and J. Carrero, “Two-photon fluorescence lifetime imaging microscopy of macrophage-mediated antigen processing,” J. Microsc. 185(3), 339–353 (1997).
[Crossref] [PubMed]

Weitz, R.

Willaert, K.

N. Boens, W. Qin, N. Basarić, J. Hofkens, M. Ameloot, J. Pouget, J.-P. Lefèvre, B. Valeur, E. Gratton, M. vandeVen, N. D. Silva, Y. Engelborghs, K. Willaert, A. Sillen, G. Rumbles, D. Phillips, A. J. Visser, A. van Hoek, J. R. Lakowicz, H. Malak, I. Gryczynski, A. G. Szabo, D. T. Krajcarski, N. Tamai, and A. Miura, “Fluorescence Lifetime Standards for Time and Frequency Domain Fluorescence Spectroscopy,” Anal. Chem. 79(5), 2137–2149 (2007).
[Crossref] [PubMed]

Williams, D. R.

R. Sharma, C. Schwarz, D. R. Williams, G. Palczewska, K. Palczewski, and J. J. Hunter, “In Vivo Two-Photon Fluorescence Kinetics of Primate Rods and Cones,” Invest. Ophthalmol. Vis. Sci. 57(2), 647–657 (2016).
[Crossref] [PubMed]

R. Sharma, D. R. Williams, G. Palczewska, K. Palczewski, and J. J. Hunter, “Two-Photon Autofluorescence Imaging Reveals Cellular Structures Throughout the Retina of the Living Primate Eye,” Invest. Ophthalmol. Vis. Sci. 57(2), 632–646 (2016).
[Crossref] [PubMed]

L. Yin, B. Masella, D. Dalkara, J. Zhang, J. G. Flannery, D. V. Schaffer, D. R. Williams, and W. H. Merigan, “Imaging Light Responses of Foveal Ganglion Cells in the Living Macaque Eye,” J. Neurosci. 34(19), 6596–6605 (2014).
[Crossref] [PubMed]

G. Palczewska, Z. Dong, M. Golczak, J. J. Hunter, D. R. Williams, N. S. Alexander, and K. Palczewski, “Noninvasive two-photon microscopy imaging of mouse retina and retinal pigment epithelium through the pupil of the eye,” Nat. Med. 20(7), 785–789 (2014).
[Crossref] [PubMed]

Q. Yang, J. Zhang, K. Nozato, K. Saito, D. R. Williams, A. Roorda, and E. A. Rossi, “Closed-loop optical stabilization and digital image registration in adaptive optics scanning light ophthalmoscopy,” Biomed. Opt. Express 5(9), 3174–3191 (2014).
[Crossref] [PubMed]

R. Sharma, L. Yin, Y. Geng, W. H. Merigan, G. Palczewska, K. Palczewski, D. R. Williams, and J. J. Hunter, “In vivo two-photon imaging of the mouse retina,” Biomed. Opt. Express 4(8), 1285–1293 (2013).
[Crossref] [PubMed]

L. Yin, Y. Geng, F. Osakada, R. Sharma, A. H. Cetin, E. M. Callaway, D. R. Williams, and W. H. Merigan, “Imaging light responses of retinal ganglion cells in the living mouse eye,” J. Neurophysiol. 109(9), 2415–2421 (2013).
[Crossref] [PubMed]

E. A. Rossi, P. Rangel-Fonseca, K. Parkins, W. Fischer, L. R. Latchney, M. A. Folwell, D. R. Williams, A. Dubra, and M. M. Chung, “In vivo imaging of retinal pigment epithelium cells in age related macular degeneration,” Biomed. Opt. Express 4(11), 2527–2539 (2013).
[Crossref] [PubMed]

Y. Geng, A. Dubra, L. Yin, W. H. Merigan, R. Sharma, R. T. Libby, and D. R. Williams, “Adaptive optics retinal imaging in the living mouse eye,” Biomed. Opt. Express 3(4), 715–734 (2012).
[Crossref] [PubMed]

Y. Geng, L. A. Schery, R. Sharma, A. Dubra, K. Ahmad, R. T. Libby, and D. R. Williams, “Optical properties of the mouse eye,” Biomed. Opt. Express 2(4), 717–738 (2011).
[Crossref] [PubMed]

J. J. Hunter, B. Masella, A. Dubra, R. Sharma, L. Yin, W. H. Merigan, G. Palczewska, K. Palczewski, and D. R. Williams, “Images of photoreceptors in living primate eyes using adaptive optics two-photon ophthalmoscopy,” Biomed. Opt. Express 2(1), 139–148 (2010).
[Crossref] [PubMed]

J. I. W. Morgan, A. Dubra, R. Wolfe, W. H. Merigan, and D. R. Williams, “In Vivo Autofluorescence Imaging of the Human and Macaque Retinal Pigment Epithelial Cell Mosaic,” Invest. Ophthalmol. Vis. Sci. 50(3), 1350–1359 (2009).
[Crossref] [PubMed]

Wolf, S.

C. Dysli, S. Wolf, K. Hatz, and M. S. Zinkernagel, “Fluorescence Lifetime Imaging in Stargardt Disease: Potential Marker for Disease Progression,” Invest. Ophthalmol. Vis. Sci. 57(3), 832–841 (2016).
[Crossref] [PubMed]

C. Dysli, M. Dysli, V. Enzmann, S. Wolf, and M. S. Zinkernagel, “Fluorescence Lifetime Imaging of the Ocular Fundus in Mice,” Invest. Ophthalmol. Vis. Sci. 55(11), 7206–7215 (2014).
[Crossref] [PubMed]

Wolfe, R.

J. I. W. Morgan, A. Dubra, R. Wolfe, W. H. Merigan, and D. R. Williams, “In Vivo Autofluorescence Imaging of the Human and Macaque Retinal Pigment Epithelial Cell Mosaic,” Invest. Ophthalmol. Vis. Sci. 50(3), 1350–1359 (2009).
[Crossref] [PubMed]

Wong, R.

D. Magde, R. Wong, and P. G. Seybold, “Fluorescence Quantum Yields and Their Relation to Lifetimes of Rhodamine 6G and Fluorescein in Nine Solvents: Improved Absolute Standards for Quantum Yields,” Photochem. Photobiol. 75(4), 327–334 (2002).
[Crossref] [PubMed]

Woods, R. L.

F. Delori, J. P. Greenberg, R. L. Woods, J. Fischer, T. Duncker, J. Sparrow, and R. T. Smith, “Quantitative measurements of autofluorescence with the scanning laser ophthalmoscope,” Invest. Ophthalmol. Vis. Sci. 52(13), 9379–9390 (2011).
[Crossref] [PubMed]

Wotzlaw, C.

E. Spiess, F. Bestvater, A. Heckel-Pompey, K. Toth, M. Hacker, G. Stobrawa, T. Feurer, C. Wotzlaw, U. Berchner-Pfannschmidt, T. Porwol, and H. Acker, “Two-photon excitation and emission spectra of the green fluorescent protein variants ECFP, EGFP and EYFP,” J. Microsc. 217(3), 200–204 (2005).
[Crossref] [PubMed]

Yang, Q.

Yeh, S.-C. A.

Yellen, G.

M. Tantama, Y. P. Hung, and G. Yellen, “Imaging intracellular pH in live cells with a genetically encoded red fluorescent protein sensor,” J. Am. Chem. Soc. 133(26), 10034–10037 (2011).
[Crossref] [PubMed]

Yin, L.

Zawadzki, R. J.

Zhang, J.

L. Yin, B. Masella, D. Dalkara, J. Zhang, J. G. Flannery, D. V. Schaffer, D. R. Williams, and W. H. Merigan, “Imaging Light Responses of Foveal Ganglion Cells in the Living Macaque Eye,” J. Neurosci. 34(19), 6596–6605 (2014).
[Crossref] [PubMed]

Q. Yang, J. Zhang, K. Nozato, K. Saito, D. R. Williams, A. Roorda, and E. A. Rossi, “Closed-loop optical stabilization and digital image registration in adaptive optics scanning light ophthalmoscopy,” Biomed. Opt. Express 5(9), 3174–3191 (2014).
[Crossref] [PubMed]

Zheng, L.

J. Dillon, L. Zheng, J. C. Merriam, and E. R. Gaillard, “Transmission spectra of light to the mammalian retina,” Photochem. Photobiol. 71(2), 225–229 (2000).
[Crossref] [PubMed]

Zinkernagel, M. S.

C. Dysli, S. Wolf, K. Hatz, and M. S. Zinkernagel, “Fluorescence Lifetime Imaging in Stargardt Disease: Potential Marker for Disease Progression,” Invest. Ophthalmol. Vis. Sci. 57(3), 832–841 (2016).
[Crossref] [PubMed]

C. Dysli, M. Dysli, V. Enzmann, S. Wolf, and M. S. Zinkernagel, “Fluorescence Lifetime Imaging of the Ocular Fundus in Mice,” Invest. Ophthalmol. Vis. Sci. 55(11), 7206–7215 (2014).
[Crossref] [PubMed]

Anal. Biochem. (1)

J. R. Lakowicz, J. Malicka, S. D’Auria, and I. Gryczynski, “Release of the self-quenching of fluorescence near silver metallic surfaces,” Anal. Biochem. 320(1), 13–20 (2003).
[Crossref] [PubMed]

Anal. Chem. (1)

N. Boens, W. Qin, N. Basarić, J. Hofkens, M. Ameloot, J. Pouget, J.-P. Lefèvre, B. Valeur, E. Gratton, M. vandeVen, N. D. Silva, Y. Engelborghs, K. Willaert, A. Sillen, G. Rumbles, D. Phillips, A. J. Visser, A. van Hoek, J. R. Lakowicz, H. Malak, I. Gryczynski, A. G. Szabo, D. T. Krajcarski, N. Tamai, and A. Miura, “Fluorescence Lifetime Standards for Time and Frequency Domain Fluorescence Spectroscopy,” Anal. Chem. 79(5), 2137–2149 (2007).
[Crossref] [PubMed]

Biomed. Opt. Express (10)

J. J. Hunter, B. Masella, A. Dubra, R. Sharma, L. Yin, W. H. Merigan, G. Palczewska, K. Palczewski, and D. R. Williams, “Images of photoreceptors in living primate eyes using adaptive optics two-photon ophthalmoscopy,” Biomed. Opt. Express 2(1), 139–148 (2010).
[Crossref] [PubMed]

Y. Geng, L. A. Schery, R. Sharma, A. Dubra, K. Ahmad, R. T. Libby, and D. R. Williams, “Optical properties of the mouse eye,” Biomed. Opt. Express 2(4), 717–738 (2011).
[Crossref] [PubMed]

R. W. K. Leung, S.-C. A. Yeh, and Q. Fang, “Effects of incomplete decay in fluorescence lifetime estimation,” Biomed. Opt. Express 2(9), 2517–2531 (2011).
[Crossref] [PubMed]

Y. Geng, A. Dubra, L. Yin, W. H. Merigan, R. Sharma, R. T. Libby, and D. R. Williams, “Adaptive optics retinal imaging in the living mouse eye,” Biomed. Opt. Express 3(4), 715–734 (2012).
[Crossref] [PubMed]

R. Sharma, L. Yin, Y. Geng, W. H. Merigan, G. Palczewska, K. Palczewski, D. R. Williams, and J. J. Hunter, “In vivo two-photon imaging of the mouse retina,” Biomed. Opt. Express 4(8), 1285–1293 (2013).
[Crossref] [PubMed]

A. Pinhas, M. Dubow, N. Shah, T. Y. Chui, D. Scoles, Y. N. Sulai, R. Weitz, J. B. Walsh, J. Carroll, A. Dubra, and R. B. Rosen, “In vivo imaging of human retinal microvasculature using adaptive optics scanning light ophthalmoscope fluorescein angiography,” Biomed. Opt. Express 4(8), 1305–1317 (2013).
[Crossref] [PubMed]

E. A. Rossi, P. Rangel-Fonseca, K. Parkins, W. Fischer, L. R. Latchney, M. A. Folwell, D. R. Williams, A. Dubra, and M. M. Chung, “In vivo imaging of retinal pigment epithelium cells in age related macular degeneration,” Biomed. Opt. Express 4(11), 2527–2539 (2013).
[Crossref] [PubMed]

Q. Yang, J. Zhang, K. Nozato, K. Saito, D. R. Williams, A. Roorda, and E. A. Rossi, “Closed-loop optical stabilization and digital image registration in adaptive optics scanning light ophthalmoscopy,” Biomed. Opt. Express 5(9), 3174–3191 (2014).
[Crossref] [PubMed]

D. J. Wahl, Y. Jian, S. Bonora, R. J. Zawadzki, and M. V. Sarunic, “Wavefront sensorless adaptive optics fluorescence biomicroscope for in vivo retinal imaging in mice,” Biomed. Opt. Express 7(1), 1–12 (2015).
[Crossref] [PubMed]

A. J. Walsh, J. T. Sharick, M. C. Skala, and H. T. Beier, “Temporal binning of time-correlated single photon counting data improves exponential decay fits and imaging speed,” Biomed. Opt. Express 7(4), 1385–1399 (2016).
[Crossref] [PubMed]

Biophys. J. (2)

C. Chen, E. Tsina, M. C. Cornwall, R. K. Crouch, S. Vijayaraghavan, and Y. Koutalos, “Reduction of all-Trans Retinal to All-Trans Retinol in the Outer Segments of Frog and Mouse Rod Photoreceptors,” Biophys. J. 88(3), 2278–2287 (2005).
[Crossref] [PubMed]

S. T. Hess, E. D. Sheets, A. Wagenknecht-Wiesner, and A. A. Heikal, “Quantitative Analysis of the Fluorescence Properties of Intrinsically Fluorescent Proteins in Living Cells,” Biophys. J. 85(4), 2566–2580 (2003).
[Crossref] [PubMed]

Chem. Rev. (1)

M. Y. Berezin and S. Achilefu, “Fluorescence Lifetime Measurements and Biological Imaging,” Chem. Rev. 110(5), 2641–2684 (2010).
[Crossref] [PubMed]

Curr. Biol. (1)

R. Pepperkok, A. Squire, S. Geley, and P. I. H. Bastiaens, “Simultaneous detection of multiple green fluorescent proteins in live cells by fluorescence lifetime imaging microscopy,” Curr. Biol. 9(5), 269–272 (1999).
[Crossref] [PubMed]

Invest. Ophthalmol. Vis. Sci. (7)

C. Dysli, M. Dysli, V. Enzmann, S. Wolf, and M. S. Zinkernagel, “Fluorescence Lifetime Imaging of the Ocular Fundus in Mice,” Invest. Ophthalmol. Vis. Sci. 55(11), 7206–7215 (2014).
[Crossref] [PubMed]

C. Dysli, S. Wolf, K. Hatz, and M. S. Zinkernagel, “Fluorescence Lifetime Imaging in Stargardt Disease: Potential Marker for Disease Progression,” Invest. Ophthalmol. Vis. Sci. 57(3), 832–841 (2016).
[Crossref] [PubMed]

J. Tam, J. Liu, A. Dubra, and R. Fariss, “In Vivo Imaging of the Human Retinal Pigment Epithelial Mosaic Using Adaptive Optics Enhanced Indocyanine Green Ophthalmoscopy,” Invest. Ophthalmol. Vis. Sci. 57(10), 4376–4384 (2016).
[Crossref] [PubMed]

J. I. W. Morgan, A. Dubra, R. Wolfe, W. H. Merigan, and D. R. Williams, “In Vivo Autofluorescence Imaging of the Human and Macaque Retinal Pigment Epithelial Cell Mosaic,” Invest. Ophthalmol. Vis. Sci. 50(3), 1350–1359 (2009).
[Crossref] [PubMed]

R. Sharma, D. R. Williams, G. Palczewska, K. Palczewski, and J. J. Hunter, “Two-Photon Autofluorescence Imaging Reveals Cellular Structures Throughout the Retina of the Living Primate Eye,” Invest. Ophthalmol. Vis. Sci. 57(2), 632–646 (2016).
[Crossref] [PubMed]

R. Sharma, C. Schwarz, D. R. Williams, G. Palczewska, K. Palczewski, and J. J. Hunter, “In Vivo Two-Photon Fluorescence Kinetics of Primate Rods and Cones,” Invest. Ophthalmol. Vis. Sci. 57(2), 647–657 (2016).
[Crossref] [PubMed]

F. Delori, J. P. Greenberg, R. L. Woods, J. Fischer, T. Duncker, J. Sparrow, and R. T. Smith, “Quantitative measurements of autofluorescence with the scanning laser ophthalmoscope,” Invest. Ophthalmol. Vis. Sci. 52(13), 9379–9390 (2011).
[Crossref] [PubMed]

J. Am. Chem. Soc. (1)

M. Tantama, Y. P. Hung, and G. Yellen, “Imaging intracellular pH in live cells with a genetically encoded red fluorescent protein sensor,” J. Am. Chem. Soc. 133(26), 10034–10037 (2011).
[Crossref] [PubMed]

J. Biomed. Opt. (2)

D. Schweitzer, M. Hammer, F. Schweitzer, R. Anders, T. Doebbecke, S. Schenke, E. R. Gaillard, and E. R. Gaillard, “In vivo measurement of time-resolved autofluorescence at the human fundus,” J. Biomed. Opt. 9(6), 1214–1222 (2004).
[Crossref] [PubMed]

D. Schweitzer, L. Deutsch, M. Klemm, S. Jentsch, M. Hammer, S. Peters, J. Haueisen, U. A. Müller, and J. Dawczynski, “Fluorescence lifetime imaging ophthalmoscopy in type 2 diabetic patients who have no signs of diabetic retinopathy,” J. Biomed. Opt. 20(6), 61106 (2015).
[Crossref] [PubMed]

J. Microsc. (2)

E. Spiess, F. Bestvater, A. Heckel-Pompey, K. Toth, M. Hacker, G. Stobrawa, T. Feurer, C. Wotzlaw, U. Berchner-Pfannschmidt, T. Porwol, and H. Acker, “Two-photon excitation and emission spectra of the green fluorescent protein variants ECFP, EGFP and EYFP,” J. Microsc. 217(3), 200–204 (2005).
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Figures (6)

Fig. 1
Fig. 1 System diagram of two-photon AOSLO designed for the mouse eye. PMT: Photomultiplier tube. SHWS: Shack-Hartmann wavefront sensor. 80/20: 80/20 beam splitter. EF: Emission filters.

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Fig. 2
Fig. 2 Example histograms and fits for (a) fluorescein in vasculature, and (b) an EGFP-labeled cell. The red open circles correspond to the number of photons collected for each time bin. The blue dashed line shows the double-exponential fit to the data. The black solid line is the IRF of the system, which is de-convolved from the data. This fit was performed at each pixel within the image which met the threshold criteria (see section 3.2).

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Fig. 3
Fig. 3 Binning analysis of the EGFP-labeled cell from Fig. 5. Panel (a) shows the two-photon fluorescence intensity image of the cell. Panels (b), (c), and (d) show fluorescence lifetime images of the cell with no binning, 3 x 3 binning, and 5 x 5 binning, respectively. A very low threshold of photons was set in order to show the inaccuracy of the fluorescence lifetime calculation when too few photons are present in the decay. The black outline in panels (b), (c) and (d) indicates the boundaries of the cell, with pixels inside the outline containing greater than 2000 photons with 5 x 5 binning. Panel (e) shows histograms of the lifetime calculated at each pixel with the different binning factors shown in (b), (c), and (d). As the binning factor increases, the lifetime fit converges on the expected lifetime of EGFP. Scale bar is 5 µm.

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Fig. 4
Fig. 4 Images of a capillary bed in the mouse inner retina. Panel (a) is a two-photon fluorescence intensity image acquired with a standard, analog PMT. Panel (b) is an image of a sub-portion of the location from panel (a) acquired with the single photon counting detector and the TCSPC system. The pixel density in this image is reduced compared with panel (a), however the structure of the capillaries is still visible. Panel (c) is a fluorescence lifetime image of the same location as panel (b). The fluorescence lifetime image is mostly uniform because the fluorescence lifetime is robust against intensity variations. Scale bar is 25 µm.

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Fig. 5
Fig. 5 Images of an EGFP-labeled ganglion cell in a Thy1-EGFP mouse retina. Panel (a) shows sparse EGFP labeling of cells and axons in the mouse retina. The blue box denotes the region of AOFLIO imaging which is shown in panel (b). Panel (b) is a two-photon fluorescence intensity image of a ganglion cell soma. The same fluorescent axon can be seen in panels (a) and (b). Panel (c) is a fluorescence lifetime image of the same location. The lifetime calculated was consistent throughout the soma of the cell, despite the intensity of fluorescence changing throughout. The fluorescence lifetime was not calculated in the axon because it did not reach threshold criteria. Scale bar is 25 µm.

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Fig. 6
Fig. 6 - Images of an EGFP-labeled cell surrounded by a bed of capillaries with fluorescein. Panel (a) shows the wide field image of the ganglion cell and its axon connecting to the optic nerve. Panel (b) shows the overlapping emission spectra for EGFP (purple dashed line) and fluorescein (red solid line), acquired from Chroma Technology Corp. Panel (c) shows the two-photon fluorescence intensity image; the cell is indistinguishable from the vessels. Panel (d) shows the fluorescence lifetime image. The EGFP fluorescence from the cell exhibits a lower fluorescence lifetime than the fluorescein, revealing the location of the cell. Panels (e) and (f) are intensity images which have been scaled by a1(x,y) (the contribution of the fast lifetime component) or a2(x,y) (the slow lifetime component), respectively. The fast lifetime component corresponds to EGFP fluorescence, causing the cell to appear in (e), while the slow lifetime component corresponds to the fluorescein, suppressing the cell in (f) and improving the vessel contrast. Scale bar is 100 µm for panel (a) and 15 µm for panels (c) – (f).

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Tables (1)

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Table 1 Fluorescence Lifetime Values measured with AOFLIO

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Equations (3)

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I( t )=ex p − t τ +C
I( t )=  a 1 ex p − t τ 1 + a 2 ex p − t τ 2 +C
τ m = a 1 τ 1 + a 2 τ 2

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