Abstract

An instrument is demonstrated that is capable of three-dimensional (3D) vasculature imaging and pO2 quantification with high spatial resolution. The instrument combines two-photon (2P) microscopy with phosphorescence quenching to measure pO2. The instrument was demonstrated by performing depth-resolved microvascular pO2 measurements of rat cortical vessels down to 120μm below the surface. 2P excitation of porphyrin was confirmed, and measured pO2 values were consistent with previously published data for normoxic and hyperoxic conditions. The ability to perform 3D pO2 measurements using optical techniques will allow researchers to overcome existing limitations imposed by polarographic electrodes, magnetic resonance techniques, and surface-only pO2 measurement techniques.

© 2008 Optical Society of America

Full Article  |  PDF Article

References

  • View by:
  • |
  • |
  • |

  1. J. M. Vanderkooi and D. F. Wilson, Adv. Exp. Med. Biol. 200, 189 (1986).
    [PubMed]
  2. J. M. Vanderkooi, G. Maniara, and T. J. Green, J. Biol. Chem. 262, 5476 (1987).
    [PubMed]
  3. R. P. Briñas, T. Troxler, and R. M. Hochstrasser, J. Am. Chem. Soc. 127, 11851 (2005).
    [CrossRef] [PubMed]
  4. M. Shibata, S. Ichioka, and J. Ando, J. Appl. Physiol. 91, 321 (2001).
    [PubMed]
  5. R. D. Shonat and A. C. Kight, Ann. Biomed. Eng. 31, 1084 (2003).
    [CrossRef] [PubMed]
  6. L. Zheng, A. S. Golub, and R. N. Pittman, Am. J. Physiol. 271, H365 (1996).
    [PubMed]
  7. R. D. Shonat, K. N. Richmond, and P. C. Johnson, Rev. Sci. Instrum. 66, 5075 (1995).
    [CrossRef]
  8. E. G. Mik, T. G. V. Leeuwen, and N. J. Raat, J. Appl. Physiol. 97, 1962 (2004).
    [CrossRef] [PubMed]
  9. D. Kleinfeld and W. Denk, in Imaging Neurons (Cold Spring Harbor Laboratory Press, 1999).
  10. I. Dunphy, S. A. Vinogradov, and D. F. Wilson, Anal. Biochem. 310, 191 (2002).
    [CrossRef] [PubMed]
  11. R. D. Shonat, E. S. Wachman, and W. Niu, Biophys. J. 73, 1223 (1997).
    [CrossRef] [PubMed]

2005 (1)

R. P. Briñas, T. Troxler, and R. M. Hochstrasser, J. Am. Chem. Soc. 127, 11851 (2005).
[CrossRef] [PubMed]

2004 (1)

E. G. Mik, T. G. V. Leeuwen, and N. J. Raat, J. Appl. Physiol. 97, 1962 (2004).
[CrossRef] [PubMed]

2003 (1)

R. D. Shonat and A. C. Kight, Ann. Biomed. Eng. 31, 1084 (2003).
[CrossRef] [PubMed]

2002 (1)

I. Dunphy, S. A. Vinogradov, and D. F. Wilson, Anal. Biochem. 310, 191 (2002).
[CrossRef] [PubMed]

2001 (1)

M. Shibata, S. Ichioka, and J. Ando, J. Appl. Physiol. 91, 321 (2001).
[PubMed]

1999 (1)

D. Kleinfeld and W. Denk, in Imaging Neurons (Cold Spring Harbor Laboratory Press, 1999).

1997 (1)

R. D. Shonat, E. S. Wachman, and W. Niu, Biophys. J. 73, 1223 (1997).
[CrossRef] [PubMed]

1996 (1)

L. Zheng, A. S. Golub, and R. N. Pittman, Am. J. Physiol. 271, H365 (1996).
[PubMed]

1995 (1)

R. D. Shonat, K. N. Richmond, and P. C. Johnson, Rev. Sci. Instrum. 66, 5075 (1995).
[CrossRef]

1987 (1)

J. M. Vanderkooi, G. Maniara, and T. J. Green, J. Biol. Chem. 262, 5476 (1987).
[PubMed]

1986 (1)

J. M. Vanderkooi and D. F. Wilson, Adv. Exp. Med. Biol. 200, 189 (1986).
[PubMed]

Ando, J.

M. Shibata, S. Ichioka, and J. Ando, J. Appl. Physiol. 91, 321 (2001).
[PubMed]

Briñas, R. P.

R. P. Briñas, T. Troxler, and R. M. Hochstrasser, J. Am. Chem. Soc. 127, 11851 (2005).
[CrossRef] [PubMed]

Denk, W.

D. Kleinfeld and W. Denk, in Imaging Neurons (Cold Spring Harbor Laboratory Press, 1999).

Dunphy, I.

I. Dunphy, S. A. Vinogradov, and D. F. Wilson, Anal. Biochem. 310, 191 (2002).
[CrossRef] [PubMed]

Golub, A. S.

L. Zheng, A. S. Golub, and R. N. Pittman, Am. J. Physiol. 271, H365 (1996).
[PubMed]

Green, T. J.

J. M. Vanderkooi, G. Maniara, and T. J. Green, J. Biol. Chem. 262, 5476 (1987).
[PubMed]

Hochstrasser, R. M.

R. P. Briñas, T. Troxler, and R. M. Hochstrasser, J. Am. Chem. Soc. 127, 11851 (2005).
[CrossRef] [PubMed]

Ichioka, S.

M. Shibata, S. Ichioka, and J. Ando, J. Appl. Physiol. 91, 321 (2001).
[PubMed]

Johnson, P. C.

R. D. Shonat, K. N. Richmond, and P. C. Johnson, Rev. Sci. Instrum. 66, 5075 (1995).
[CrossRef]

Kight, A. C.

R. D. Shonat and A. C. Kight, Ann. Biomed. Eng. 31, 1084 (2003).
[CrossRef] [PubMed]

Kleinfeld, D.

D. Kleinfeld and W. Denk, in Imaging Neurons (Cold Spring Harbor Laboratory Press, 1999).

Leeuwen, T. G. V.

E. G. Mik, T. G. V. Leeuwen, and N. J. Raat, J. Appl. Physiol. 97, 1962 (2004).
[CrossRef] [PubMed]

Maniara, G.

J. M. Vanderkooi, G. Maniara, and T. J. Green, J. Biol. Chem. 262, 5476 (1987).
[PubMed]

Mik, E. G.

E. G. Mik, T. G. V. Leeuwen, and N. J. Raat, J. Appl. Physiol. 97, 1962 (2004).
[CrossRef] [PubMed]

Niu, W.

R. D. Shonat, E. S. Wachman, and W. Niu, Biophys. J. 73, 1223 (1997).
[CrossRef] [PubMed]

Pittman, R. N.

L. Zheng, A. S. Golub, and R. N. Pittman, Am. J. Physiol. 271, H365 (1996).
[PubMed]

Raat, N. J.

E. G. Mik, T. G. V. Leeuwen, and N. J. Raat, J. Appl. Physiol. 97, 1962 (2004).
[CrossRef] [PubMed]

Richmond, K. N.

R. D. Shonat, K. N. Richmond, and P. C. Johnson, Rev. Sci. Instrum. 66, 5075 (1995).
[CrossRef]

Shibata, M.

M. Shibata, S. Ichioka, and J. Ando, J. Appl. Physiol. 91, 321 (2001).
[PubMed]

Shonat, R. D.

R. D. Shonat and A. C. Kight, Ann. Biomed. Eng. 31, 1084 (2003).
[CrossRef] [PubMed]

R. D. Shonat, E. S. Wachman, and W. Niu, Biophys. J. 73, 1223 (1997).
[CrossRef] [PubMed]

R. D. Shonat, K. N. Richmond, and P. C. Johnson, Rev. Sci. Instrum. 66, 5075 (1995).
[CrossRef]

Troxler, T.

R. P. Briñas, T. Troxler, and R. M. Hochstrasser, J. Am. Chem. Soc. 127, 11851 (2005).
[CrossRef] [PubMed]

Vanderkooi, J. M.

J. M. Vanderkooi, G. Maniara, and T. J. Green, J. Biol. Chem. 262, 5476 (1987).
[PubMed]

J. M. Vanderkooi and D. F. Wilson, Adv. Exp. Med. Biol. 200, 189 (1986).
[PubMed]

Vinogradov, S. A.

I. Dunphy, S. A. Vinogradov, and D. F. Wilson, Anal. Biochem. 310, 191 (2002).
[CrossRef] [PubMed]

Wachman, E. S.

R. D. Shonat, E. S. Wachman, and W. Niu, Biophys. J. 73, 1223 (1997).
[CrossRef] [PubMed]

Wilson, D. F.

I. Dunphy, S. A. Vinogradov, and D. F. Wilson, Anal. Biochem. 310, 191 (2002).
[CrossRef] [PubMed]

J. M. Vanderkooi and D. F. Wilson, Adv. Exp. Med. Biol. 200, 189 (1986).
[PubMed]

Zheng, L.

L. Zheng, A. S. Golub, and R. N. Pittman, Am. J. Physiol. 271, H365 (1996).
[PubMed]

Adv. Exp. Med. Biol. (1)

J. M. Vanderkooi and D. F. Wilson, Adv. Exp. Med. Biol. 200, 189 (1986).
[PubMed]

Am. J. Physiol. (1)

L. Zheng, A. S. Golub, and R. N. Pittman, Am. J. Physiol. 271, H365 (1996).
[PubMed]

Anal. Biochem. (1)

I. Dunphy, S. A. Vinogradov, and D. F. Wilson, Anal. Biochem. 310, 191 (2002).
[CrossRef] [PubMed]

Ann. Biomed. Eng. (1)

R. D. Shonat and A. C. Kight, Ann. Biomed. Eng. 31, 1084 (2003).
[CrossRef] [PubMed]

Biophys. J. (1)

R. D. Shonat, E. S. Wachman, and W. Niu, Biophys. J. 73, 1223 (1997).
[CrossRef] [PubMed]

J. Am. Chem. Soc. (1)

R. P. Briñas, T. Troxler, and R. M. Hochstrasser, J. Am. Chem. Soc. 127, 11851 (2005).
[CrossRef] [PubMed]

J. Appl. Physiol. (2)

M. Shibata, S. Ichioka, and J. Ando, J. Appl. Physiol. 91, 321 (2001).
[PubMed]

E. G. Mik, T. G. V. Leeuwen, and N. J. Raat, J. Appl. Physiol. 97, 1962 (2004).
[CrossRef] [PubMed]

J. Biol. Chem. (1)

J. M. Vanderkooi, G. Maniara, and T. J. Green, J. Biol. Chem. 262, 5476 (1987).
[PubMed]

Rev. Sci. Instrum. (1)

R. D. Shonat, K. N. Richmond, and P. C. Johnson, Rev. Sci. Instrum. 66, 5075 (1995).
[CrossRef]

Other (1)

D. Kleinfeld and W. Denk, in Imaging Neurons (Cold Spring Harbor Laboratory Press, 1999).

Cited By

OSA participates in CrossRef's Cited-By Linking service. Citing articles from OSA journals and other participating publishers are listed here.

Alert me when this article is cited.


Figures (3)

Fig. 1
Fig. 1

Instrumentation setup. AOM is added to a standard 2P microscope setup to facilitate pO 2 measurement.

Fig. 2
Fig. 2

Image of arteriole network 120 μ m below the surface under (a) normoxia and (b) hyperoxia. The phosphorescence decay data with best fit curves for the highlighted point in panels (a) and (b) are depicted in (c). Image of arteriole network 100 μ m below the surface under (d) normoxia and (e) hyperoxia. The phosphorescence decay data with best fit curves for the highlighted point in panels (d) and (e) are depicted in (f). pO 2 values are in units of mmHg. (g) Verification of 2P excitation of porphyrin.

Fig. 3
Fig. 3

Image of branching cortical vessel 60 μ m below the surface. A clear pO 2 gradient is visible owing to an occlusion in the lower branch of the vessel. pO 2 values are in units of mmHg.

Metrics