Abstract

The flow of individual corpuscles through retinal capillaries may now be observed noninvasively by using adaptive optics (AO). To explore their imaging properties, we imaged retinal capillary flow in two healthy subjects at 593 nm with a flood-based AO ophthalmoscope, at a variety of retinal locations and levels of defocus. The image intensity of red cells and plasma depends upon capillary depth relative to focus: red cells appear brighter than background, and plasma darker, for capillaries posterior to focus. The reverse is true for capillaries anterior to focus. Contrast reversals were obtained over 0.05 D (14μm), which are well within the typical undulations in depth of retinal capillaries. We relate these observations to phase-contrast defocusing microscopy. This defocusing effect confounds flow measurements, which rely on correlation of image intensity between successive locations along the same capillary, a requirement made further difficult by high physiological variability in flow. Peak correlation was maintained >0.25 over a distance of 22±15μm (roughly the spacing between red cells) and over a duration of 154±49ms (roughly eight times the temporal period between red cells). We provide a 2D correlogram approach that significantly improves robustness in the face of optical and physiological variability, compared to the traditional spatiotemporal plot, without requiring additional data.

© 2014 Optical Society of America

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References

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  1. J. Liang and D. R. Williams, J. Opt. Soc. Am. A 14, 2873 (1997).
    [CrossRef]
  2. Z. Zhong, B. L. Petrig, X. Qi, and S. A. Burns, Opt. Express 16, 12746 (2008).
    [CrossRef]
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    [CrossRef]
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    [CrossRef]
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    [CrossRef]
  6. J. Tam, P. Tiruveedhula, and A. Roorda, Biomed. Opt. Express 2, 781 (2011).
    [CrossRef]
  7. A. Uji, M. Hangai, S. Ooto, K. Takayama, N. Arakawa, H. Imamura, K. Nozato, and N. Yoshimura, Invest. Ophthalmol. Vis. Sci. 53, 171 (2012).
    [CrossRef]
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    [CrossRef]
  9. B. P. Helmke, S. N. Bremner, B. W. Zweifach, R. Skalak, and G. W. Schmid-Schonbein, Am. J. Physiol. 273, H2884 (1997).
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    [CrossRef]
  11. H. H. Lipowsky, N. U. Sheikh, and D. M. Katz, J. Clin. Invest. 80, 117 (1987).
    [CrossRef]
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    [CrossRef]
  13. U. Agero, C. H. Monken, C. Ropert, R. T. Gazzinelli, and O. N. Mesquita, Phys. Rev. E 67, 051904 (2003).
    [CrossRef]
  14. L. G. Mesquita, U. Agero, and O. N. Mesquita, Appl. Phys. Lett. 88, 133901 (2006).
    [CrossRef]
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    [CrossRef]
  16. Y. L. Jin, J. Y. Chen, L. Xu, and P. N. Wang, Phys. Med. Biol. 51, N371 (2006).
    [CrossRef]
  17. A. Ajo, Acta Physiologica Scandinavica 13, 130 (1947).
    [CrossRef]
  18. P. Bedggood and A. Metha, “Oximetry imaging of the retinal microvasculature using adaptive optics,” presented at the Association for Research in Vision and Ophthalmology Annual Meeting, Ft. Lauderdale, Fla., May6–10, 2012.
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    [CrossRef]
  20. B. Stefanovic, E. Hutchinson, V. Yakovleva, V. Schram, J. T. Russell, L. Belluscio, A. P. Koretsky, and A. C. Silva, J. Cereb. Blood Flow Metab. 28, 961 (2008).
    [CrossRef]
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    [CrossRef]

2012 (3)

A. Uji, M. Hangai, S. Ooto, K. Takayama, N. Arakawa, H. Imamura, K. Nozato, and N. Yoshimura, Invest. Ophthalmol. Vis. Sci. 53, 171 (2012).
[CrossRef]

Z. Zhong, G. Huang, T. Y. Chui, B. L. Petrig, and S. A. Burns, J. Vis. 12(6), 3 (2012).
[CrossRef]

P. Bedggood and A. Metha, Biomed. Opt. Express 3, 3264 (2012).
[CrossRef]

2011 (2)

J. Tam, P. Tiruveedhula, and A. Roorda, Biomed. Opt. Express 2, 781 (2011).
[CrossRef]

Z. Zhong, H. Song, T. Y. Chui, B. L. Petrig, and S. A. Burns, Invest. Ophthalmol. Vis. Sci. 52, 4151 (2011).
[CrossRef]

2010 (1)

J. Tam, J. A. Martin, and A. Roorda, Investig. Ophthalmol. Vis. Sci. 51, 1691 (2010).
[CrossRef]

2009 (1)

J. A. Martin and A. Roorda, Exp. Eye Res. 88, 356 (2009).
[CrossRef]

2008 (2)

B. Stefanovic, E. Hutchinson, V. Yakovleva, V. Schram, J. T. Russell, L. Belluscio, A. P. Koretsky, and A. C. Silva, J. Cereb. Blood Flow Metab. 28, 961 (2008).
[CrossRef]

Z. Zhong, B. L. Petrig, X. Qi, and S. A. Burns, Opt. Express 16, 12746 (2008).
[CrossRef]

2006 (2)

L. G. Mesquita, U. Agero, and O. N. Mesquita, Appl. Phys. Lett. 88, 133901 (2006).
[CrossRef]

Y. L. Jin, J. Y. Chen, L. Xu, and P. N. Wang, Phys. Med. Biol. 51, N371 (2006).
[CrossRef]

2005 (1)

J. A. Martin and A. Roorda, Ophthalmology 112, 2219 (2005).
[CrossRef]

2003 (2)

U. Agero, C. H. Monken, C. Ropert, R. T. Gazzinelli, and O. N. Mesquita, Phys. Rev. E 67, 051904 (2003).
[CrossRef]

E. Chaigneau, M. Oheim, E. Audinat, and S. Charpak, Proc. Natl. Acad. Sci. USA 100, 13081 (2003).
[CrossRef]

1999 (1)

A. Roggan, M. Friebel, K. Do Rschel, A. Hahn, and G. Mu Ller, J. Biomed. Opt. 4, 36 (1999).
[CrossRef]

1997 (2)

J. Liang and D. R. Williams, J. Opt. Soc. Am. A 14, 2873 (1997).
[CrossRef]

B. P. Helmke, S. N. Bremner, B. W. Zweifach, R. Skalak, and G. W. Schmid-Schonbein, Am. J. Physiol. 273, H2884 (1997).

1991 (1)

P. Gasser and O. Meienberg, Eur. Neurol. 31, 168 (1991).
[CrossRef]

1987 (1)

H. H. Lipowsky, N. U. Sheikh, and D. M. Katz, J. Clin. Invest. 80, 117 (1987).
[CrossRef]

1947 (1)

A. Ajo, Acta Physiologica Scandinavica 13, 130 (1947).
[CrossRef]

Agero, U.

L. G. Mesquita, U. Agero, and O. N. Mesquita, Appl. Phys. Lett. 88, 133901 (2006).
[CrossRef]

U. Agero, C. H. Monken, C. Ropert, R. T. Gazzinelli, and O. N. Mesquita, Phys. Rev. E 67, 051904 (2003).
[CrossRef]

Ajo, A.

A. Ajo, Acta Physiologica Scandinavica 13, 130 (1947).
[CrossRef]

Arakawa, N.

A. Uji, M. Hangai, S. Ooto, K. Takayama, N. Arakawa, H. Imamura, K. Nozato, and N. Yoshimura, Invest. Ophthalmol. Vis. Sci. 53, 171 (2012).
[CrossRef]

Audinat, E.

E. Chaigneau, M. Oheim, E. Audinat, and S. Charpak, Proc. Natl. Acad. Sci. USA 100, 13081 (2003).
[CrossRef]

Bedggood, P.

P. Bedggood and A. Metha, Biomed. Opt. Express 3, 3264 (2012).
[CrossRef]

P. Bedggood and A. Metha, “Oximetry imaging of the retinal microvasculature using adaptive optics,” presented at the Association for Research in Vision and Ophthalmology Annual Meeting, Ft. Lauderdale, Fla., May6–10, 2012.

Belluscio, L.

B. Stefanovic, E. Hutchinson, V. Yakovleva, V. Schram, J. T. Russell, L. Belluscio, A. P. Koretsky, and A. C. Silva, J. Cereb. Blood Flow Metab. 28, 961 (2008).
[CrossRef]

Bremner, S. N.

B. P. Helmke, S. N. Bremner, B. W. Zweifach, R. Skalak, and G. W. Schmid-Schonbein, Am. J. Physiol. 273, H2884 (1997).

Burns, S. A.

Z. Zhong, G. Huang, T. Y. Chui, B. L. Petrig, and S. A. Burns, J. Vis. 12(6), 3 (2012).
[CrossRef]

Z. Zhong, H. Song, T. Y. Chui, B. L. Petrig, and S. A. Burns, Invest. Ophthalmol. Vis. Sci. 52, 4151 (2011).
[CrossRef]

Z. Zhong, B. L. Petrig, X. Qi, and S. A. Burns, Opt. Express 16, 12746 (2008).
[CrossRef]

Chaigneau, E.

E. Chaigneau, M. Oheim, E. Audinat, and S. Charpak, Proc. Natl. Acad. Sci. USA 100, 13081 (2003).
[CrossRef]

Charpak, S.

E. Chaigneau, M. Oheim, E. Audinat, and S. Charpak, Proc. Natl. Acad. Sci. USA 100, 13081 (2003).
[CrossRef]

Chen, J. Y.

Y. L. Jin, J. Y. Chen, L. Xu, and P. N. Wang, Phys. Med. Biol. 51, N371 (2006).
[CrossRef]

Chui, T. Y.

Z. Zhong, G. Huang, T. Y. Chui, B. L. Petrig, and S. A. Burns, J. Vis. 12(6), 3 (2012).
[CrossRef]

Z. Zhong, H. Song, T. Y. Chui, B. L. Petrig, and S. A. Burns, Invest. Ophthalmol. Vis. Sci. 52, 4151 (2011).
[CrossRef]

Do Rschel, K.

A. Roggan, M. Friebel, K. Do Rschel, A. Hahn, and G. Mu Ller, J. Biomed. Opt. 4, 36 (1999).
[CrossRef]

Friebel, M.

A. Roggan, M. Friebel, K. Do Rschel, A. Hahn, and G. Mu Ller, J. Biomed. Opt. 4, 36 (1999).
[CrossRef]

Gasser, P.

P. Gasser and O. Meienberg, Eur. Neurol. 31, 168 (1991).
[CrossRef]

Gazzinelli, R. T.

U. Agero, C. H. Monken, C. Ropert, R. T. Gazzinelli, and O. N. Mesquita, Phys. Rev. E 67, 051904 (2003).
[CrossRef]

Hahn, A.

A. Roggan, M. Friebel, K. Do Rschel, A. Hahn, and G. Mu Ller, J. Biomed. Opt. 4, 36 (1999).
[CrossRef]

Hangai, M.

A. Uji, M. Hangai, S. Ooto, K. Takayama, N. Arakawa, H. Imamura, K. Nozato, and N. Yoshimura, Invest. Ophthalmol. Vis. Sci. 53, 171 (2012).
[CrossRef]

Helmke, B. P.

B. P. Helmke, S. N. Bremner, B. W. Zweifach, R. Skalak, and G. W. Schmid-Schonbein, Am. J. Physiol. 273, H2884 (1997).

Huang, G.

Z. Zhong, G. Huang, T. Y. Chui, B. L. Petrig, and S. A. Burns, J. Vis. 12(6), 3 (2012).
[CrossRef]

Hutchinson, E.

B. Stefanovic, E. Hutchinson, V. Yakovleva, V. Schram, J. T. Russell, L. Belluscio, A. P. Koretsky, and A. C. Silva, J. Cereb. Blood Flow Metab. 28, 961 (2008).
[CrossRef]

Imamura, H.

A. Uji, M. Hangai, S. Ooto, K. Takayama, N. Arakawa, H. Imamura, K. Nozato, and N. Yoshimura, Invest. Ophthalmol. Vis. Sci. 53, 171 (2012).
[CrossRef]

Jin, Y. L.

Y. L. Jin, J. Y. Chen, L. Xu, and P. N. Wang, Phys. Med. Biol. 51, N371 (2006).
[CrossRef]

Katz, D. M.

H. H. Lipowsky, N. U. Sheikh, and D. M. Katz, J. Clin. Invest. 80, 117 (1987).
[CrossRef]

Koretsky, A. P.

B. Stefanovic, E. Hutchinson, V. Yakovleva, V. Schram, J. T. Russell, L. Belluscio, A. P. Koretsky, and A. C. Silva, J. Cereb. Blood Flow Metab. 28, 961 (2008).
[CrossRef]

Liang, J.

Lipowsky, H. H.

H. H. Lipowsky, N. U. Sheikh, and D. M. Katz, J. Clin. Invest. 80, 117 (1987).
[CrossRef]

Martin, J. A.

J. Tam, J. A. Martin, and A. Roorda, Investig. Ophthalmol. Vis. Sci. 51, 1691 (2010).
[CrossRef]

J. A. Martin and A. Roorda, Exp. Eye Res. 88, 356 (2009).
[CrossRef]

J. A. Martin and A. Roorda, Ophthalmology 112, 2219 (2005).
[CrossRef]

Meienberg, O.

P. Gasser and O. Meienberg, Eur. Neurol. 31, 168 (1991).
[CrossRef]

Mesquita, L. G.

L. G. Mesquita, U. Agero, and O. N. Mesquita, Appl. Phys. Lett. 88, 133901 (2006).
[CrossRef]

Mesquita, O. N.

L. G. Mesquita, U. Agero, and O. N. Mesquita, Appl. Phys. Lett. 88, 133901 (2006).
[CrossRef]

U. Agero, C. H. Monken, C. Ropert, R. T. Gazzinelli, and O. N. Mesquita, Phys. Rev. E 67, 051904 (2003).
[CrossRef]

Metha, A.

P. Bedggood and A. Metha, Biomed. Opt. Express 3, 3264 (2012).
[CrossRef]

P. Bedggood and A. Metha, “Oximetry imaging of the retinal microvasculature using adaptive optics,” presented at the Association for Research in Vision and Ophthalmology Annual Meeting, Ft. Lauderdale, Fla., May6–10, 2012.

Monken, C. H.

U. Agero, C. H. Monken, C. Ropert, R. T. Gazzinelli, and O. N. Mesquita, Phys. Rev. E 67, 051904 (2003).
[CrossRef]

Mu Ller, G.

A. Roggan, M. Friebel, K. Do Rschel, A. Hahn, and G. Mu Ller, J. Biomed. Opt. 4, 36 (1999).
[CrossRef]

Nozato, K.

A. Uji, M. Hangai, S. Ooto, K. Takayama, N. Arakawa, H. Imamura, K. Nozato, and N. Yoshimura, Invest. Ophthalmol. Vis. Sci. 53, 171 (2012).
[CrossRef]

Oheim, M.

E. Chaigneau, M. Oheim, E. Audinat, and S. Charpak, Proc. Natl. Acad. Sci. USA 100, 13081 (2003).
[CrossRef]

Ooto, S.

A. Uji, M. Hangai, S. Ooto, K. Takayama, N. Arakawa, H. Imamura, K. Nozato, and N. Yoshimura, Invest. Ophthalmol. Vis. Sci. 53, 171 (2012).
[CrossRef]

Petrig, B. L.

Z. Zhong, G. Huang, T. Y. Chui, B. L. Petrig, and S. A. Burns, J. Vis. 12(6), 3 (2012).
[CrossRef]

Z. Zhong, H. Song, T. Y. Chui, B. L. Petrig, and S. A. Burns, Invest. Ophthalmol. Vis. Sci. 52, 4151 (2011).
[CrossRef]

Z. Zhong, B. L. Petrig, X. Qi, and S. A. Burns, Opt. Express 16, 12746 (2008).
[CrossRef]

Qi, X.

Roggan, A.

A. Roggan, M. Friebel, K. Do Rschel, A. Hahn, and G. Mu Ller, J. Biomed. Opt. 4, 36 (1999).
[CrossRef]

Roorda, A.

J. Tam, P. Tiruveedhula, and A. Roorda, Biomed. Opt. Express 2, 781 (2011).
[CrossRef]

J. Tam, J. A. Martin, and A. Roorda, Investig. Ophthalmol. Vis. Sci. 51, 1691 (2010).
[CrossRef]

J. A. Martin and A. Roorda, Exp. Eye Res. 88, 356 (2009).
[CrossRef]

J. A. Martin and A. Roorda, Ophthalmology 112, 2219 (2005).
[CrossRef]

Ropert, C.

U. Agero, C. H. Monken, C. Ropert, R. T. Gazzinelli, and O. N. Mesquita, Phys. Rev. E 67, 051904 (2003).
[CrossRef]

Russell, J. T.

B. Stefanovic, E. Hutchinson, V. Yakovleva, V. Schram, J. T. Russell, L. Belluscio, A. P. Koretsky, and A. C. Silva, J. Cereb. Blood Flow Metab. 28, 961 (2008).
[CrossRef]

Schmid-Schonbein, G. W.

B. P. Helmke, S. N. Bremner, B. W. Zweifach, R. Skalak, and G. W. Schmid-Schonbein, Am. J. Physiol. 273, H2884 (1997).

Schram, V.

B. Stefanovic, E. Hutchinson, V. Yakovleva, V. Schram, J. T. Russell, L. Belluscio, A. P. Koretsky, and A. C. Silva, J. Cereb. Blood Flow Metab. 28, 961 (2008).
[CrossRef]

Sheikh, N. U.

H. H. Lipowsky, N. U. Sheikh, and D. M. Katz, J. Clin. Invest. 80, 117 (1987).
[CrossRef]

Silva, A. C.

B. Stefanovic, E. Hutchinson, V. Yakovleva, V. Schram, J. T. Russell, L. Belluscio, A. P. Koretsky, and A. C. Silva, J. Cereb. Blood Flow Metab. 28, 961 (2008).
[CrossRef]

Skalak, R.

B. P. Helmke, S. N. Bremner, B. W. Zweifach, R. Skalak, and G. W. Schmid-Schonbein, Am. J. Physiol. 273, H2884 (1997).

Song, H.

Z. Zhong, H. Song, T. Y. Chui, B. L. Petrig, and S. A. Burns, Invest. Ophthalmol. Vis. Sci. 52, 4151 (2011).
[CrossRef]

Stefanovic, B.

B. Stefanovic, E. Hutchinson, V. Yakovleva, V. Schram, J. T. Russell, L. Belluscio, A. P. Koretsky, and A. C. Silva, J. Cereb. Blood Flow Metab. 28, 961 (2008).
[CrossRef]

Takayama, K.

A. Uji, M. Hangai, S. Ooto, K. Takayama, N. Arakawa, H. Imamura, K. Nozato, and N. Yoshimura, Invest. Ophthalmol. Vis. Sci. 53, 171 (2012).
[CrossRef]

Tam, J.

J. Tam, P. Tiruveedhula, and A. Roorda, Biomed. Opt. Express 2, 781 (2011).
[CrossRef]

J. Tam, J. A. Martin, and A. Roorda, Investig. Ophthalmol. Vis. Sci. 51, 1691 (2010).
[CrossRef]

Tiruveedhula, P.

Uji, A.

A. Uji, M. Hangai, S. Ooto, K. Takayama, N. Arakawa, H. Imamura, K. Nozato, and N. Yoshimura, Invest. Ophthalmol. Vis. Sci. 53, 171 (2012).
[CrossRef]

Wang, P. N.

Y. L. Jin, J. Y. Chen, L. Xu, and P. N. Wang, Phys. Med. Biol. 51, N371 (2006).
[CrossRef]

Williams, D. R.

Xu, L.

Y. L. Jin, J. Y. Chen, L. Xu, and P. N. Wang, Phys. Med. Biol. 51, N371 (2006).
[CrossRef]

Yakovleva, V.

B. Stefanovic, E. Hutchinson, V. Yakovleva, V. Schram, J. T. Russell, L. Belluscio, A. P. Koretsky, and A. C. Silva, J. Cereb. Blood Flow Metab. 28, 961 (2008).
[CrossRef]

Yoshimura, N.

A. Uji, M. Hangai, S. Ooto, K. Takayama, N. Arakawa, H. Imamura, K. Nozato, and N. Yoshimura, Invest. Ophthalmol. Vis. Sci. 53, 171 (2012).
[CrossRef]

Zhong, Z.

Z. Zhong, G. Huang, T. Y. Chui, B. L. Petrig, and S. A. Burns, J. Vis. 12(6), 3 (2012).
[CrossRef]

Z. Zhong, H. Song, T. Y. Chui, B. L. Petrig, and S. A. Burns, Invest. Ophthalmol. Vis. Sci. 52, 4151 (2011).
[CrossRef]

Z. Zhong, B. L. Petrig, X. Qi, and S. A. Burns, Opt. Express 16, 12746 (2008).
[CrossRef]

Zweifach, B. W.

B. P. Helmke, S. N. Bremner, B. W. Zweifach, R. Skalak, and G. W. Schmid-Schonbein, Am. J. Physiol. 273, H2884 (1997).

Acta Physiologica Scandinavica (1)

A. Ajo, Acta Physiologica Scandinavica 13, 130 (1947).
[CrossRef]

Am. J. Physiol. (1)

B. P. Helmke, S. N. Bremner, B. W. Zweifach, R. Skalak, and G. W. Schmid-Schonbein, Am. J. Physiol. 273, H2884 (1997).

Appl. Phys. Lett. (1)

L. G. Mesquita, U. Agero, and O. N. Mesquita, Appl. Phys. Lett. 88, 133901 (2006).
[CrossRef]

Biomed. Opt. Express (2)

Eur. Neurol. (1)

P. Gasser and O. Meienberg, Eur. Neurol. 31, 168 (1991).
[CrossRef]

Exp. Eye Res. (1)

J. A. Martin and A. Roorda, Exp. Eye Res. 88, 356 (2009).
[CrossRef]

Invest. Ophthalmol. Vis. Sci. (2)

A. Uji, M. Hangai, S. Ooto, K. Takayama, N. Arakawa, H. Imamura, K. Nozato, and N. Yoshimura, Invest. Ophthalmol. Vis. Sci. 53, 171 (2012).
[CrossRef]

Z. Zhong, H. Song, T. Y. Chui, B. L. Petrig, and S. A. Burns, Invest. Ophthalmol. Vis. Sci. 52, 4151 (2011).
[CrossRef]

Investig. Ophthalmol. Vis. Sci. (1)

J. Tam, J. A. Martin, and A. Roorda, Investig. Ophthalmol. Vis. Sci. 51, 1691 (2010).
[CrossRef]

J. Biomed. Opt. (1)

A. Roggan, M. Friebel, K. Do Rschel, A. Hahn, and G. Mu Ller, J. Biomed. Opt. 4, 36 (1999).
[CrossRef]

J. Cereb. Blood Flow Metab. (1)

B. Stefanovic, E. Hutchinson, V. Yakovleva, V. Schram, J. T. Russell, L. Belluscio, A. P. Koretsky, and A. C. Silva, J. Cereb. Blood Flow Metab. 28, 961 (2008).
[CrossRef]

J. Clin. Invest. (1)

H. H. Lipowsky, N. U. Sheikh, and D. M. Katz, J. Clin. Invest. 80, 117 (1987).
[CrossRef]

J. Opt. Soc. Am. A (1)

J. Vis. (1)

Z. Zhong, G. Huang, T. Y. Chui, B. L. Petrig, and S. A. Burns, J. Vis. 12(6), 3 (2012).
[CrossRef]

Ophthalmology (1)

J. A. Martin and A. Roorda, Ophthalmology 112, 2219 (2005).
[CrossRef]

Opt. Express (1)

Phys. Med. Biol. (1)

Y. L. Jin, J. Y. Chen, L. Xu, and P. N. Wang, Phys. Med. Biol. 51, N371 (2006).
[CrossRef]

Phys. Rev. E (1)

U. Agero, C. H. Monken, C. Ropert, R. T. Gazzinelli, and O. N. Mesquita, Phys. Rev. E 67, 051904 (2003).
[CrossRef]

Proc. Natl. Acad. Sci. USA (1)

E. Chaigneau, M. Oheim, E. Audinat, and S. Charpak, Proc. Natl. Acad. Sci. USA 100, 13081 (2003).
[CrossRef]

Other (1)

P. Bedggood and A. Metha, “Oximetry imaging of the retinal microvasculature using adaptive optics,” presented at the Association for Research in Vision and Ophthalmology Annual Meeting, Ft. Lauderdale, Fla., May6–10, 2012.

Supplementary Material (2)

» Media 1: AVI (2105 KB)     
» Media 2: AVI (2466 KB)     

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Figures (4)

Fig. 1.
Fig. 1.

Intensity of corpuscles in capillary segments as a function of defocus. Black: dark corpuscles; white: bright corpuscles; gray: ambiguous corpuscles (e.g., due to loss of contrast). Bars indicate proportion of vessel pixels in each state (black, gray, or white). Representative sequences shown in Media 1 (PB, 2° inferior, 1° nasal) and Media 2 (JZ, 1° superior, 1° temporal). Scale: each panel is 410×190μm.

Fig. 2.
Fig. 2.

Spatiotemporal analysis in one vessel: (a) spatiotemporal plot and (b) 2D spatial correlogram. Arrows indicate longest distance over which peak of correlation with central point remained above threshold. (c) 2D temporal correlogram. (d) Radon transform of (a). (e) Variance of the radon transform for the spatiotemporal plot (blue), 2D spatial correlogram (red), and 2D temporal correlogram (black).

Fig. 3.
Fig. 3.

Coherence of flow data over space (left) and over time (right). Data averaged over 171 vessel segments. Error bars show ± one standard deviation from the mean.

Fig. 4.
Fig. 4.

Spatiotemporal plot (left) and the variance in its radon transform (right) for two different vessel segments (top and bottom). Straight lines indicate slope determined from three complementary approaches: the spatiotemporal plot (blue), the 2D spatial correlogram (red), or the 2D temporal correlogram (black). For vessels in which subjective confirmation of slope was deemed reliable, strong agreement was always obtained by accepting the alternative with greatest “peakedness” (ratio of peak/median for the variance of the Radon transform). This corresponded to one of the 2D correlograms in 87% of vessel segments analyzed.

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