Abstract

Scanning-laser ophthalmoscope (SLO) technology has provided, among other possibilities, the potential for three-dimensional (3-D) visualization of anatomy in the posterior pole of the eye. The use of indocyanine green (ICG) as an infrared fluorescent marker of vasculature in combination with an infrared SLO (the Heidelberg Retina Angiograph) is presented. Presently, two main factors among others discussed impede the visualization of 3-D structures in observed SLO data. Random eye motion between optical sections and (to a lesser degree) motion between raster scan lines prevent assessment of spatial orientation and connectivity of vasculature. Second, smear along the optic axis owing to the optics prevents accurate determination of vessel or lesion size and shape, especially for features spanning several optical sections. A novel, to our knowledge, deconvolution algorithm is described that automatically corrects for the poor axial (optical-sectioning) resolution of the SLO and for patient random eye motion during target fixation. Encouraging preliminary results are presented showing the usefulness of applying blind deconvolution toward improving the 3-D clarity of SLO data. Although clinical and medical research applications are broad, the specific medical sample selected shows the potential of examining microvascular 3-D morphology for diagnosis and treatment of choroidal tumors.

© 1998 Optical Society of America

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1995 (3)

D. U. Bartsch, R. N. Weinreb, G. Zinser, W. R. Freeman, “Confocal scanning infrared laser ophthalmoscopy for indocyanine green angiography: preliminary results,” Am. J. Ophthalmol. 120, 642–651 (1995).
[PubMed]

A. V. Cideciyan, “Registration of ocular fundus images,” IEEE Eng. Med. Biol. 14, 52–58 (1995).
[CrossRef]

V. Krishnamurti, Y. Liu, S. Bhattacharyya, J. N. Turner, T. J. Holmes, “Blind deconvolution of fluorescence micrographs by maximum-likelihood estimation,” Appl. Opt. 34, 6633–6647 (1995).
[CrossRef]

1994 (5)

V. Rummelt, R. Folberg, C. Rummelt, L. Gruman, T. Hwang, R. Woolson, M. Hong Yi, G. Naumann, “Microcirculation architecture of melanocytic nevi and malignant melanomas of the ciliary body and choroid,” Ophthalmology 101, 718–727 (1994).
[PubMed]

Q. Chen, M. Defrise, F. Deconinck, “Symmetric phase-only matched filtering of Fourier–Mellin transforms for image registration and recognition,” IEEE Trans. Pattern Anal. Mach. Intell. 16, 1156–1168 (1994).
[CrossRef]

D. R. Guyer, L. A. Yannuzzi, J. S. Slakter, J. A. Sorenson, A. Ho, D. Orlock, “Digital indocyanine green videoangiography of central serous chorioretinopathy,” Arch. Ophthalmol. 112, 1057–1062 (1994).
[CrossRef] [PubMed]

U. Introini, A. Mosca, R. Brancato, A. Pece, R. Paleari, “Spectrofluorimetric analysis of indocyanine green fluorescence in patients with age-related macular degeneration,” (abstract) Invest. Ophthalmol. Vis. Sci. (Suppl.) 35, 1500 (1994).

A. C. Ho, L. A. Yannuzzi, D. R. Guyer, J. S. Slakter, J. A. Sorenson, D. A. Orlock, “Intraretinal leakage of indocyanine green dye,” Ophthalmology 101, 534–541 (1994).
[PubMed]

1993 (3)

Collaborative Ocular Melanoma Study Group, “The Collaborative Ocular Melanoma Study: COMS report no. 3,” Contr. Clin. Trials 14, 362–391 (1993).

R. Folberg, V. Rummelt, R. Parys-Van Ginderdeuren, T. Hwang, R. Woolson, J. Peer, L. Gruman, “The prognostic value of tumor blood vessel morphology in primary uveal melanoma,” Ophthalmology 100, 1389–1398 (1993).
[PubMed]

S. Joshi, M. I. Miller, “Maximum a posteriori estimation with Good’s roughness for three-dimensional optical sectioning microscopy,” J. Opt. Soc. Am. A 10, 1078–1085 (1993).
[CrossRef] [PubMed]

1992 (2)

L. G. Brown, “A survey of image registration techniques,” ACM Comput. Surv. 24 (4), 325–376 (1992).
[CrossRef]

T. J. Holmes, “Blind deconvolution of quantum-limited incoherent imagery,” J. Opt. Soc. Am. A 9, 1052–1061 (1992).
[CrossRef] [PubMed]

1991 (3)

M. I. Miller, B. Roysam, “Bayesian image reconstruction for emission tomography incorporating Good’s roughness prior on massively parallel processors,” Proc. Natl. Acad. Sci. 88, 3223–3227 (1991).
[CrossRef]

T. J. Holmes, Y. Liu, “Acceleration of maximum-likelihood image restoration for fluorescence microscopy and other noncoherent imagery,” J. Opt. Soc. Am. A 8, 893–907 (1991).
[CrossRef]

S. Wolf, O. Arend, H. Toonen, “Retinal capillary blood flow measurement with scanning laser ophthalmoscope. Preliminary results,” Ophthalmology 98, 996–1000 (1991).
[PubMed]

1989 (3)

D. U. Bartsch, M. Intaglietta, J. F. Bille, A. W. Dreher, M. Gharib, W. R. Freeman, “Confocal laser tomographic analysis of the retina in eyes with macular hole formation and other focal macular diseases,” Am. J. Ophthalmol. 108, 277–287 (1989).
[PubMed]

A. Scheider, C. Schroedel, “High resolution indocyanine green angiography with a scanning laser ophthalmoscope,” Am. J. Ophthalmol. 108, 458–459 (1989).
[PubMed]

R. A. Robb, C. Barillot, “Interactive display and analysis of 3D medical images,” IEEE Trans. Med. Imaging 8, 217–226 (1989).
[CrossRef]

1988 (2)

1987 (3)

R. H. Webb, G. W. Hughes, F. C. Delori, “Confocal scanning laser ophthalmoscope,” Appl. Opt. 26, 1492–1499 (1987).
[CrossRef] [PubMed]

E. DeCastro, G. Cristini, A. Martelli, C. Morandi, M. Vascotto, “Compensation of random eye motion in television ophthalmoscopy: preliminary results,” IEEE Trans. Med. Imaging 6, 74–81 (1987).
[CrossRef]

E. DeCastro, C. Morandi, “Registration of translated and rotated images using finite Fourier transforms,” IEEE Trans. Pattern Anal. Mach. Intell. 9, 700–703 (1987).
[CrossRef]

1982 (1)

L. A. Shepp, Y. Vardi, “Maximum likelihood reconstruction for emission tomography,” IEEE Trans. Med. Imaging MI-1, 113–122 (1982).
[CrossRef]

1980 (3)

R. H. Webb, G. W. Hughes, O. Pomerantzeff, “Flying spot TV ophthalmoscope,” Appl. Opt. 19, 2991–2997 (1980).
[CrossRef] [PubMed]

I. McLean, W. Foster, L. Zimmerman, “Choroidal melanoma,” Arch. Ophthalmol. 98, 1298–1303 (1980).
[CrossRef] [PubMed]

J. Shields, J. Augsburger, G. Brown, R. Stephens, “The differential diagnosis of posterior uveal melanoma,” Ophthalmology 87, 518–522 (1980).
[PubMed]

1978 (1)

L. Zimmerman, I. McLean, W. Foster, “Does enucleation of the eye containing a malignant melanoma prevent or accelerate the dissemination of tumour cells,” Br. J. Ophthalmol. 62, 420–425 (1978).
[CrossRef] [PubMed]

1977 (1)

A. P. Dempster, N. M. Laird, D. B. Rubin, “Maximum likelihood from incomplete data via the EM algorithm,” J. R. Stat. Soc. B 39, 1–37 (1977).

1975 (1)

K. Ossoinig, F. Bigar, S. Kaefring, “Malignant melanoma of the choroid and ciliary body. A differential diagnosis in clinical echography,” Bibl. Ophthalmol. 83, 141–154 (1975).
[PubMed]

1974 (1)

G. W. Gerchberg, W. O. Saxton, “Super-resolution through error energy reduction,” Opt. Acta 21, 709–720 (1974).
[CrossRef]

1973 (1)

R. W. Flower, B. F. Hochheimer, “A clinical technique and apparatus for simultaneous angiography of the separate retinal and choroidal circulations,” Invest. Ophthalmol. 12, 248–261 (1973).
[PubMed]

1971 (1)

B. F. Hochheimer, “Angiography of the retina with indocyanine green,” Arch. Ophthalmol. 86, 564–565 (1971).
[CrossRef] [PubMed]

1966 (1)

F. W. Campbell, R. W. Gubisch, “Optical quality of the human eye,” J. Physiol. 186, 558–578 (1966).
[PubMed]

Arend, O.

S. Wolf, O. Arend, H. Toonen, “Retinal capillary blood flow measurement with scanning laser ophthalmoscope. Preliminary results,” Ophthalmology 98, 996–1000 (1991).
[PubMed]

Augsburger, J.

J. Shields, J. Augsburger, G. Brown, R. Stephens, “The differential diagnosis of posterior uveal melanoma,” Ophthalmology 87, 518–522 (1980).
[PubMed]

Ayers, G. R.

Barillot, C.

R. A. Robb, C. Barillot, “Interactive display and analysis of 3D medical images,” IEEE Trans. Med. Imaging 8, 217–226 (1989).
[CrossRef]

Bartsch, D. U.

D. U. Bartsch, R. N. Weinreb, G. Zinser, W. R. Freeman, “Confocal scanning infrared laser ophthalmoscopy for indocyanine green angiography: preliminary results,” Am. J. Ophthalmol. 120, 642–651 (1995).
[PubMed]

D. U. Bartsch, M. Intaglietta, J. F. Bille, A. W. Dreher, M. Gharib, W. R. Freeman, “Confocal laser tomographic analysis of the retina in eyes with macular hole formation and other focal macular diseases,” Am. J. Ophthalmol. 108, 277–287 (1989).
[PubMed]

A. J. Mueller, D. U. Bartsch, R Folberg, J. Peer, M. H. Goldbaum, D. G. Bitner, R. N. Weinreb, W. R. Freeman, “Imaging the microvascularization of choroidal melanomas using confocal indocyanine green scanning laser ophthalmoscopy,” Arch. Ophthalmol. (to be published).

Bhattacharyya, S.

V. Krishnamurti, Y. Liu, S. Bhattacharyya, J. N. Turner, T. J. Holmes, “Blind deconvolution of fluorescence micrographs by maximum-likelihood estimation,” Appl. Opt. 34, 6633–6647 (1995).
[CrossRef]

T. J. Holmes, S. Bhattacharyya, J. A. Cooper, D. Hanzel, V. Krishnamurthi, W. Lin, B. Roysam, D. H. Szarowski, J. N. Turner, “Light microscopic images reconstructed by maximum likelihood deconvolution,” in Handbook of Biological Confocal Microscopy, J. B. Pawley, ed. (Plenum, New York, 1995), pp. 389–402.
[CrossRef]

T. J. Holmes, S. Bhattacharyya, J. A. Cooper, D. Hanzel, V. Krishnamurthi, W. Lin, B. Roysam, D. H. Szarowski, J. N. Turner, “Simultaneous image and point spread function reconstruction for 3D light microscopy,” in Image Reconstruction and Restoration, T. J. Schulz, D. L. Snyder, eds., Proc. SPIE2302, 359–368 (1994).
[CrossRef]

S. Bhattacharyya, D. H. Szarowski, J. N. Turner, N. O’Connor, T. J. Holmes, “The ML–blind deconvolution algorithm: recent developments,” in Three-Dimensional Microscopy: Image Acquisition and Processing III, C. J. Cogswell, G. S. Kino, T. Wilson, eds., Proc. SPIE2655, 175–186 (1996).
[CrossRef]

Bigar, F.

K. Ossoinig, F. Bigar, S. Kaefring, “Malignant melanoma of the choroid and ciliary body. A differential diagnosis in clinical echography,” Bibl. Ophthalmol. 83, 141–154 (1975).
[PubMed]

Bille, J. F.

D. U. Bartsch, M. Intaglietta, J. F. Bille, A. W. Dreher, M. Gharib, W. R. Freeman, “Confocal laser tomographic analysis of the retina in eyes with macular hole formation and other focal macular diseases,” Am. J. Ophthalmol. 108, 277–287 (1989).
[PubMed]

Bitner, D. G.

A. J. Mueller, D. U. Bartsch, R Folberg, J. Peer, M. H. Goldbaum, D. G. Bitner, R. N. Weinreb, W. R. Freeman, “Imaging the microvascularization of choroidal melanomas using confocal indocyanine green scanning laser ophthalmoscopy,” Arch. Ophthalmol. (to be published).

Brancato, R.

U. Introini, A. Mosca, R. Brancato, A. Pece, R. Paleari, “Spectrofluorimetric analysis of indocyanine green fluorescence in patients with age-related macular degeneration,” (abstract) Invest. Ophthalmol. Vis. Sci. (Suppl.) 35, 1500 (1994).

Brown, G.

J. Shields, J. Augsburger, G. Brown, R. Stephens, “The differential diagnosis of posterior uveal melanoma,” Ophthalmology 87, 518–522 (1980).
[PubMed]

Brown, L. G.

L. G. Brown, “A survey of image registration techniques,” ACM Comput. Surv. 24 (4), 325–376 (1992).
[CrossRef]

Campbell, F. W.

F. W. Campbell, R. W. Gubisch, “Optical quality of the human eye,” J. Physiol. 186, 558–578 (1966).
[PubMed]

Chen, Q.

Q. Chen, M. Defrise, F. Deconinck, “Symmetric phase-only matched filtering of Fourier–Mellin transforms for image registration and recognition,” IEEE Trans. Pattern Anal. Mach. Intell. 16, 1156–1168 (1994).
[CrossRef]

Cideciyan, A. V.

A. V. Cideciyan, “Registration of ocular fundus images,” IEEE Eng. Med. Biol. 14, 52–58 (1995).
[CrossRef]

Cooper, J. A.

T. J. Holmes, S. Bhattacharyya, J. A. Cooper, D. Hanzel, V. Krishnamurthi, W. Lin, B. Roysam, D. H. Szarowski, J. N. Turner, “Light microscopic images reconstructed by maximum likelihood deconvolution,” in Handbook of Biological Confocal Microscopy, J. B. Pawley, ed. (Plenum, New York, 1995), pp. 389–402.
[CrossRef]

T. J. Holmes, S. Bhattacharyya, J. A. Cooper, D. Hanzel, V. Krishnamurthi, W. Lin, B. Roysam, D. H. Szarowski, J. N. Turner, “Simultaneous image and point spread function reconstruction for 3D light microscopy,” in Image Reconstruction and Restoration, T. J. Schulz, D. L. Snyder, eds., Proc. SPIE2302, 359–368 (1994).
[CrossRef]

Cristini, G.

E. DeCastro, G. Cristini, A. Martelli, C. Morandi, M. Vascotto, “Compensation of random eye motion in television ophthalmoscopy: preliminary results,” IEEE Trans. Med. Imaging 6, 74–81 (1987).
[CrossRef]

Dainty, J. C.

DeCastro, E.

E. DeCastro, C. Morandi, “Registration of translated and rotated images using finite Fourier transforms,” IEEE Trans. Pattern Anal. Mach. Intell. 9, 700–703 (1987).
[CrossRef]

E. DeCastro, G. Cristini, A. Martelli, C. Morandi, M. Vascotto, “Compensation of random eye motion in television ophthalmoscopy: preliminary results,” IEEE Trans. Med. Imaging 6, 74–81 (1987).
[CrossRef]

Deconinck, F.

Q. Chen, M. Defrise, F. Deconinck, “Symmetric phase-only matched filtering of Fourier–Mellin transforms for image registration and recognition,” IEEE Trans. Pattern Anal. Mach. Intell. 16, 1156–1168 (1994).
[CrossRef]

Defrise, M.

Q. Chen, M. Defrise, F. Deconinck, “Symmetric phase-only matched filtering of Fourier–Mellin transforms for image registration and recognition,” IEEE Trans. Pattern Anal. Mach. Intell. 16, 1156–1168 (1994).
[CrossRef]

Delori, F. C.

Dempster, A. P.

A. P. Dempster, N. M. Laird, D. B. Rubin, “Maximum likelihood from incomplete data via the EM algorithm,” J. R. Stat. Soc. B 39, 1–37 (1977).

Dreher, A. W.

D. U. Bartsch, M. Intaglietta, J. F. Bille, A. W. Dreher, M. Gharib, W. R. Freeman, “Confocal laser tomographic analysis of the retina in eyes with macular hole formation and other focal macular diseases,” Am. J. Ophthalmol. 108, 277–287 (1989).
[PubMed]

Flower, R. W.

R. W. Flower, B. F. Hochheimer, “A clinical technique and apparatus for simultaneous angiography of the separate retinal and choroidal circulations,” Invest. Ophthalmol. 12, 248–261 (1973).
[PubMed]

Folberg, R

A. J. Mueller, D. U. Bartsch, R Folberg, J. Peer, M. H. Goldbaum, D. G. Bitner, R. N. Weinreb, W. R. Freeman, “Imaging the microvascularization of choroidal melanomas using confocal indocyanine green scanning laser ophthalmoscopy,” Arch. Ophthalmol. (to be published).

Folberg, R.

V. Rummelt, R. Folberg, C. Rummelt, L. Gruman, T. Hwang, R. Woolson, M. Hong Yi, G. Naumann, “Microcirculation architecture of melanocytic nevi and malignant melanomas of the ciliary body and choroid,” Ophthalmology 101, 718–727 (1994).
[PubMed]

R. Folberg, V. Rummelt, R. Parys-Van Ginderdeuren, T. Hwang, R. Woolson, J. Peer, L. Gruman, “The prognostic value of tumor blood vessel morphology in primary uveal melanoma,” Ophthalmology 100, 1389–1398 (1993).
[PubMed]

Foos, R. Y.

B. R. Straatsma, R. Y. Foos, L. M. Spencer, “The retina-topography and clinical correlations,” in Symposium on Retina and Retinal Surgery: Transactions of the New Orleans Academy of Ophthalmology, W. D. Cockerham, ed. (Mosby, St. Louis, Mo., 1969).

Foster, W.

I. McLean, W. Foster, L. Zimmerman, “Choroidal melanoma,” Arch. Ophthalmol. 98, 1298–1303 (1980).
[CrossRef] [PubMed]

L. Zimmerman, I. McLean, W. Foster, “Does enucleation of the eye containing a malignant melanoma prevent or accelerate the dissemination of tumour cells,” Br. J. Ophthalmol. 62, 420–425 (1978).
[CrossRef] [PubMed]

Freeman, W. R.

D. U. Bartsch, R. N. Weinreb, G. Zinser, W. R. Freeman, “Confocal scanning infrared laser ophthalmoscopy for indocyanine green angiography: preliminary results,” Am. J. Ophthalmol. 120, 642–651 (1995).
[PubMed]

D. U. Bartsch, M. Intaglietta, J. F. Bille, A. W. Dreher, M. Gharib, W. R. Freeman, “Confocal laser tomographic analysis of the retina in eyes with macular hole formation and other focal macular diseases,” Am. J. Ophthalmol. 108, 277–287 (1989).
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T. J. Holmes, S. Bhattacharyya, J. A. Cooper, D. Hanzel, V. Krishnamurthi, W. Lin, B. Roysam, D. H. Szarowski, J. N. Turner, “Light microscopic images reconstructed by maximum likelihood deconvolution,” in Handbook of Biological Confocal Microscopy, J. B. Pawley, ed. (Plenum, New York, 1995), pp. 389–402.
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T. J. Holmes, S. Bhattacharyya, J. A. Cooper, D. Hanzel, V. Krishnamurthi, W. Lin, B. Roysam, D. H. Szarowski, J. N. Turner, “Simultaneous image and point spread function reconstruction for 3D light microscopy,” in Image Reconstruction and Restoration, T. J. Schulz, D. L. Snyder, eds., Proc. SPIE2302, 359–368 (1994).
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A. J. Mueller, D. U. Bartsch, R Folberg, J. Peer, M. H. Goldbaum, D. G. Bitner, R. N. Weinreb, W. R. Freeman, “Imaging the microvascularization of choroidal melanomas using confocal indocyanine green scanning laser ophthalmoscopy,” Arch. Ophthalmol. (to be published).

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V. Rummelt, R. Folberg, C. Rummelt, L. Gruman, T. Hwang, R. Woolson, M. Hong Yi, G. Naumann, “Microcirculation architecture of melanocytic nevi and malignant melanomas of the ciliary body and choroid,” Ophthalmology 101, 718–727 (1994).
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S. Bhattacharyya, D. H. Szarowski, J. N. Turner, N. O’Connor, T. J. Holmes, “The ML–blind deconvolution algorithm: recent developments,” in Three-Dimensional Microscopy: Image Acquisition and Processing III, C. J. Cogswell, G. S. Kino, T. Wilson, eds., Proc. SPIE2655, 175–186 (1996).
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D. R. Guyer, L. A. Yannuzzi, J. S. Slakter, J. A. Sorenson, A. Ho, D. Orlock, “Digital indocyanine green videoangiography of central serous chorioretinopathy,” Arch. Ophthalmol. 112, 1057–1062 (1994).
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A. C. Ho, L. A. Yannuzzi, D. R. Guyer, J. S. Slakter, J. A. Sorenson, D. A. Orlock, “Intraretinal leakage of indocyanine green dye,” Ophthalmology 101, 534–541 (1994).
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K. Ossoinig, F. Bigar, S. Kaefring, “Malignant melanoma of the choroid and ciliary body. A differential diagnosis in clinical echography,” Bibl. Ophthalmol. 83, 141–154 (1975).
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U. Introini, A. Mosca, R. Brancato, A. Pece, R. Paleari, “Spectrofluorimetric analysis of indocyanine green fluorescence in patients with age-related macular degeneration,” (abstract) Invest. Ophthalmol. Vis. Sci. (Suppl.) 35, 1500 (1994).

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R. Folberg, V. Rummelt, R. Parys-Van Ginderdeuren, T. Hwang, R. Woolson, J. Peer, L. Gruman, “The prognostic value of tumor blood vessel morphology in primary uveal melanoma,” Ophthalmology 100, 1389–1398 (1993).
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R. Folberg, V. Rummelt, R. Parys-Van Ginderdeuren, T. Hwang, R. Woolson, J. Peer, L. Gruman, “The prognostic value of tumor blood vessel morphology in primary uveal melanoma,” Ophthalmology 100, 1389–1398 (1993).
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T. J. Holmes, S. Bhattacharyya, J. A. Cooper, D. Hanzel, V. Krishnamurthi, W. Lin, B. Roysam, D. H. Szarowski, J. N. Turner, “Light microscopic images reconstructed by maximum likelihood deconvolution,” in Handbook of Biological Confocal Microscopy, J. B. Pawley, ed. (Plenum, New York, 1995), pp. 389–402.
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T. J. Holmes, S. Bhattacharyya, J. A. Cooper, D. Hanzel, V. Krishnamurthi, W. Lin, B. Roysam, D. H. Szarowski, J. N. Turner, “Simultaneous image and point spread function reconstruction for 3D light microscopy,” in Image Reconstruction and Restoration, T. J. Schulz, D. L. Snyder, eds., Proc. SPIE2302, 359–368 (1994).
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A. P. Dempster, N. M. Laird, D. B. Rubin, “Maximum likelihood from incomplete data via the EM algorithm,” J. R. Stat. Soc. B 39, 1–37 (1977).

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V. Rummelt, R. Folberg, C. Rummelt, L. Gruman, T. Hwang, R. Woolson, M. Hong Yi, G. Naumann, “Microcirculation architecture of melanocytic nevi and malignant melanomas of the ciliary body and choroid,” Ophthalmology 101, 718–727 (1994).
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V. Rummelt, R. Folberg, C. Rummelt, L. Gruman, T. Hwang, R. Woolson, M. Hong Yi, G. Naumann, “Microcirculation architecture of melanocytic nevi and malignant melanomas of the ciliary body and choroid,” Ophthalmology 101, 718–727 (1994).
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D. R. Guyer, L. A. Yannuzzi, J. S. Slakter, J. A. Sorenson, A. Ho, D. Orlock, “Digital indocyanine green videoangiography of central serous chorioretinopathy,” Arch. Ophthalmol. 112, 1057–1062 (1994).
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S. Bhattacharyya, D. H. Szarowski, J. N. Turner, N. O’Connor, T. J. Holmes, “The ML–blind deconvolution algorithm: recent developments,” in Three-Dimensional Microscopy: Image Acquisition and Processing III, C. J. Cogswell, G. S. Kino, T. Wilson, eds., Proc. SPIE2655, 175–186 (1996).
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T. J. Holmes, S. Bhattacharyya, J. A. Cooper, D. Hanzel, V. Krishnamurthi, W. Lin, B. Roysam, D. H. Szarowski, J. N. Turner, “Simultaneous image and point spread function reconstruction for 3D light microscopy,” in Image Reconstruction and Restoration, T. J. Schulz, D. L. Snyder, eds., Proc. SPIE2302, 359–368 (1994).
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T. J. Holmes, S. Bhattacharyya, J. A. Cooper, D. Hanzel, V. Krishnamurthi, W. Lin, B. Roysam, D. H. Szarowski, J. N. Turner, “Light microscopic images reconstructed by maximum likelihood deconvolution,” in Handbook of Biological Confocal Microscopy, J. B. Pawley, ed. (Plenum, New York, 1995), pp. 389–402.
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V. Krishnamurti, Y. Liu, S. Bhattacharyya, J. N. Turner, T. J. Holmes, “Blind deconvolution of fluorescence micrographs by maximum-likelihood estimation,” Appl. Opt. 34, 6633–6647 (1995).
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S. Bhattacharyya, D. H. Szarowski, J. N. Turner, N. O’Connor, T. J. Holmes, “The ML–blind deconvolution algorithm: recent developments,” in Three-Dimensional Microscopy: Image Acquisition and Processing III, C. J. Cogswell, G. S. Kino, T. Wilson, eds., Proc. SPIE2655, 175–186 (1996).
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T. J. Holmes, S. Bhattacharyya, J. A. Cooper, D. Hanzel, V. Krishnamurthi, W. Lin, B. Roysam, D. H. Szarowski, J. N. Turner, “Light microscopic images reconstructed by maximum likelihood deconvolution,” in Handbook of Biological Confocal Microscopy, J. B. Pawley, ed. (Plenum, New York, 1995), pp. 389–402.
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T. J. Holmes, S. Bhattacharyya, J. A. Cooper, D. Hanzel, V. Krishnamurthi, W. Lin, B. Roysam, D. H. Szarowski, J. N. Turner, “Simultaneous image and point spread function reconstruction for 3D light microscopy,” in Image Reconstruction and Restoration, T. J. Schulz, D. L. Snyder, eds., Proc. SPIE2302, 359–368 (1994).
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L. A. Shepp, Y. Vardi, “Maximum likelihood reconstruction for emission tomography,” IEEE Trans. Med. Imaging MI-1, 113–122 (1982).
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E. DeCastro, G. Cristini, A. Martelli, C. Morandi, M. Vascotto, “Compensation of random eye motion in television ophthalmoscopy: preliminary results,” IEEE Trans. Med. Imaging 6, 74–81 (1987).
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Webb, W. W.

D. R. Sandison, R. M. Williams, K. S. Wells, J. Strickler, W. W. Webb, “Quantitative fluorescence confocal laser scanning microscopy (CLSM),” in Handbook of Biological Confocal Microscopy, J. B. Pawley, ed. (Plenum, New York, 1995), pp. 39–53.
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D. R. Sandison, R. M. Williams, K. S. Wells, J. Strickler, W. W. Webb, “Quantitative fluorescence confocal laser scanning microscopy (CLSM),” in Handbook of Biological Confocal Microscopy, J. B. Pawley, ed. (Plenum, New York, 1995), pp. 39–53.
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D. R. Sandison, R. M. Williams, K. S. Wells, J. Strickler, W. W. Webb, “Quantitative fluorescence confocal laser scanning microscopy (CLSM),” in Handbook of Biological Confocal Microscopy, J. B. Pawley, ed. (Plenum, New York, 1995), pp. 39–53.
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S. Wolf, O. Arend, H. Toonen, “Retinal capillary blood flow measurement with scanning laser ophthalmoscope. Preliminary results,” Ophthalmology 98, 996–1000 (1991).
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V. Rummelt, R. Folberg, C. Rummelt, L. Gruman, T. Hwang, R. Woolson, M. Hong Yi, G. Naumann, “Microcirculation architecture of melanocytic nevi and malignant melanomas of the ciliary body and choroid,” Ophthalmology 101, 718–727 (1994).
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Figures (11)

Fig. 1
Fig. 1

Schematic diagram of the confocal optics of the HRA SLO.

Fig. 2
Fig. 2

Maximum-intensity projection (MIP) of a tomographic series scanned with the HRA and consisting of twenty-five 256 × 256 pixel optical sections. The series was obtained during the midphase (∼10 min after injection). Tomograms were collected with sampling spacings of 23.05 μm in the focal plane and 83.33 μm along the optic axis.

Fig. 3
Fig. 3

Cutaway view of a tomographic series demonstrating the in-plane thickness versus the axial thickness of features. Views shown are cross sections taken along the optic axis Z and along the X axis, resulting in XY (in-plane) and YZ (planar) views.

Fig. 4
Fig. 4

Flowchart of the deconvolution algorithm. See text for details.

Fig. 5
Fig. 5

Results of the coarse-registration stage. MIP’s of registered tomograms are shown. Subvolumes N and O are indicated and measure 64 × 64 pixels in X and Y.

Fig. 6
Fig. 6

Equivalent optical sections of (a) unprocessed and (b) deconvolved data taken from subvolume N.

Fig. 7
Fig. 7

Cross sections of subvolume O taken along the Y axis, producing XZ planar views. (a) XZ views of the subvolume before deconvolution and after coarse registration. (b) XZ views of the subvolume after deconvolution.

Fig. 8
Fig. 8

Optical sections from large-field deconvolution results. Equivalent adjacent optical sections of (a) coarsely registered and (b) deconvolved optical sections.

Fig. 9
Fig. 9

MIP views of (a) coarsely registered and (b) deconvolved tomograms. The axial extent of the tomographic scan is indicated at the right-hand side of (a).

Fig. 10
Fig. 10

MIP views of subfield N (a) before and (b) after deconvolution. In both (a) and (b), the XY, XZ, and YZ views are displayed from left to right. In both (a) and (b), arrows 1 and 3 indicate choroidal vessels, and arrow 2 designates a retinal vessel. Traces were performed as described in Section 4.

Fig. 11
Fig. 11

MIP views of subfield O (a) before and (b) after deconvolution. In both (a) and (b), the XY, XZ, and YZ views are displayed from left to right. The subvolume was interpolated along the optic axis before display to obtain isotropic volumetric sampling. Traces were performed as described in Section 4.

Tables (1)

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Table 1 Definitions

Equations (3)

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L = x , y , z R 3   n x ,   y ,   z log μ ( x ,   y ,   z ) d x d y d z - x , y , z R 3   μ ( x ,   y ,   z ) d x d y d z ,
L = x , y , z R 3   n x ,   y ,   z log   μ [ x + Δ x z ,   y + Δ z ,   z ] d x d y d z - x , y , z R 3   μ [ x + Δ x z ,   y + Δ y z ,   z ] d x d y d z .
L Δ x z = x , y , z R 3 1 - n x ,   y ,   z μ [ x - Δ x z ,   y - Δ y z ,   z ] ×   μ [ x - Δ x z ,   y - Δ y z ,   z ] Δ x z .

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