Abstract

We present the results of diffuse reflectance measurements made on the surface of a tissue-simulating phantom containing intact human erythrocytes. These measurements indicate that the absorption spectrum of hemoglobin in its natural environment is significantly different from that measured in homogeneous fluid solution, especially in the spectral regions of highest absorption. We show that this difference can be explained by the pigment packaging theory developed by Duysens [Biochim. Biophys. Acta 19, 1 (1956)] and that the adoption of basis spectra that take this effect into account improves the accuracy of fitting diffuse reflectance spectra.

© 2004 Optical Society of America

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References

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  1. E. M. Jope, in Hemoglobin, F. J. W. Roughton and J. C. Kendrew, eds. (Interscience, New York, 1949), pp. 205–219.
  2. O. W. Van Assendelft, Spectrophotometry of Hemoglobin Derivatives (Charles C. Thomas, Springfield, Ill., 1970).
  3. S. Takatani and M. D. Graham, IEEE Trans. Biomed. Eng. BME-25, 656 (1987).
  4. S. Wray, M. Cope, D. T. Delpy, J. S. Wyatt, and E. O. R. Reynolds, Biochim. Biophys. Acta 933, 184 (1988).
    [CrossRef] [PubMed]
  5. W. G. Zijlstra, A. Bursma, and W. P. Meeuwsen-van der Roest, Clin. Chem. 37, 1633 (1991).
    [PubMed]
  6. S. A. Prahl, “Optical properties of hemoglobin,” http://www.omlc.ogi.edu/spectra/hemoglobin/summary.html (retrieved July30, 2001).
  7. L. N. M. Duysens, Biochim. Biophys. Acta 19, 1 (1956).
    [CrossRef] [PubMed]
  8. P. Latimer and C. A. H. Eubanks, Arch. Biochem. Biophys. 98, 274 (1962).
    [CrossRef] [PubMed]
  9. G. N. Lapennas, J. M. Colacino, and J. Bonaventura, Methods Enzymol. 76, 449 (1981).
    [CrossRef]
  10. E. Loewinger, A. Gordon, A. Weinreb, and J. Gross, J. Appl. Physiol. 19, 1179 (1964).
    [PubMed]
  11. B. W. Pogue, E. A. White, U. L. Osterberg, and K. D. Paulsen, Appl. Opt. 40, 4616 (2001).
    [CrossRef]
  12. S. Patel and R. N. Pittman, Microvasc. Res. 62, 63 (2001).
    [CrossRef] [PubMed]
  13. E. L. Hull, M. G. Nichols, and T. H. Foster, Phys. Med. Biol. 43, 3381 (1998).
    [CrossRef] [PubMed]
  14. E. L. Hull and T. H. Foster, J. Opt. Soc. Am. A 18, 584 (2001).
    [CrossRef]
  15. V. Venugopalan, J. S. You, and B. J. Tromberg, Phys. Rev. E 58, 2395 (1998).
    [CrossRef]

2001 (3)

1998 (2)

V. Venugopalan, J. S. You, and B. J. Tromberg, Phys. Rev. E 58, 2395 (1998).
[CrossRef]

E. L. Hull, M. G. Nichols, and T. H. Foster, Phys. Med. Biol. 43, 3381 (1998).
[CrossRef] [PubMed]

1991 (1)

W. G. Zijlstra, A. Bursma, and W. P. Meeuwsen-van der Roest, Clin. Chem. 37, 1633 (1991).
[PubMed]

1988 (1)

S. Wray, M. Cope, D. T. Delpy, J. S. Wyatt, and E. O. R. Reynolds, Biochim. Biophys. Acta 933, 184 (1988).
[CrossRef] [PubMed]

1987 (1)

S. Takatani and M. D. Graham, IEEE Trans. Biomed. Eng. BME-25, 656 (1987).

1981 (1)

G. N. Lapennas, J. M. Colacino, and J. Bonaventura, Methods Enzymol. 76, 449 (1981).
[CrossRef]

1964 (1)

E. Loewinger, A. Gordon, A. Weinreb, and J. Gross, J. Appl. Physiol. 19, 1179 (1964).
[PubMed]

1962 (1)

P. Latimer and C. A. H. Eubanks, Arch. Biochem. Biophys. 98, 274 (1962).
[CrossRef] [PubMed]

1956 (1)

L. N. M. Duysens, Biochim. Biophys. Acta 19, 1 (1956).
[CrossRef] [PubMed]

Bonaventura, J.

G. N. Lapennas, J. M. Colacino, and J. Bonaventura, Methods Enzymol. 76, 449 (1981).
[CrossRef]

Bursma, A.

W. G. Zijlstra, A. Bursma, and W. P. Meeuwsen-van der Roest, Clin. Chem. 37, 1633 (1991).
[PubMed]

Colacino, J. M.

G. N. Lapennas, J. M. Colacino, and J. Bonaventura, Methods Enzymol. 76, 449 (1981).
[CrossRef]

Cope, M.

S. Wray, M. Cope, D. T. Delpy, J. S. Wyatt, and E. O. R. Reynolds, Biochim. Biophys. Acta 933, 184 (1988).
[CrossRef] [PubMed]

Delpy, D. T.

S. Wray, M. Cope, D. T. Delpy, J. S. Wyatt, and E. O. R. Reynolds, Biochim. Biophys. Acta 933, 184 (1988).
[CrossRef] [PubMed]

Duysens, L. N. M.

L. N. M. Duysens, Biochim. Biophys. Acta 19, 1 (1956).
[CrossRef] [PubMed]

Eubanks, C. A. H.

P. Latimer and C. A. H. Eubanks, Arch. Biochem. Biophys. 98, 274 (1962).
[CrossRef] [PubMed]

Foster, T. H.

E. L. Hull and T. H. Foster, J. Opt. Soc. Am. A 18, 584 (2001).
[CrossRef]

E. L. Hull, M. G. Nichols, and T. H. Foster, Phys. Med. Biol. 43, 3381 (1998).
[CrossRef] [PubMed]

Gordon, A.

E. Loewinger, A. Gordon, A. Weinreb, and J. Gross, J. Appl. Physiol. 19, 1179 (1964).
[PubMed]

Graham, M. D.

S. Takatani and M. D. Graham, IEEE Trans. Biomed. Eng. BME-25, 656 (1987).

Gross, J.

E. Loewinger, A. Gordon, A. Weinreb, and J. Gross, J. Appl. Physiol. 19, 1179 (1964).
[PubMed]

Hull, E. L.

E. L. Hull and T. H. Foster, J. Opt. Soc. Am. A 18, 584 (2001).
[CrossRef]

E. L. Hull, M. G. Nichols, and T. H. Foster, Phys. Med. Biol. 43, 3381 (1998).
[CrossRef] [PubMed]

Jope, E. M.

E. M. Jope, in Hemoglobin, F. J. W. Roughton and J. C. Kendrew, eds. (Interscience, New York, 1949), pp. 205–219.

Lapennas, G. N.

G. N. Lapennas, J. M. Colacino, and J. Bonaventura, Methods Enzymol. 76, 449 (1981).
[CrossRef]

Latimer, P.

P. Latimer and C. A. H. Eubanks, Arch. Biochem. Biophys. 98, 274 (1962).
[CrossRef] [PubMed]

Loewinger, E.

E. Loewinger, A. Gordon, A. Weinreb, and J. Gross, J. Appl. Physiol. 19, 1179 (1964).
[PubMed]

Meeuwsen-van der Roest, W. P.

W. G. Zijlstra, A. Bursma, and W. P. Meeuwsen-van der Roest, Clin. Chem. 37, 1633 (1991).
[PubMed]

Nichols, M. G.

E. L. Hull, M. G. Nichols, and T. H. Foster, Phys. Med. Biol. 43, 3381 (1998).
[CrossRef] [PubMed]

Osterberg, U. L.

Patel, S.

S. Patel and R. N. Pittman, Microvasc. Res. 62, 63 (2001).
[CrossRef] [PubMed]

Paulsen, K. D.

Pittman, R. N.

S. Patel and R. N. Pittman, Microvasc. Res. 62, 63 (2001).
[CrossRef] [PubMed]

Pogue, B. W.

Reynolds, E. O. R.

S. Wray, M. Cope, D. T. Delpy, J. S. Wyatt, and E. O. R. Reynolds, Biochim. Biophys. Acta 933, 184 (1988).
[CrossRef] [PubMed]

Takatani, S.

S. Takatani and M. D. Graham, IEEE Trans. Biomed. Eng. BME-25, 656 (1987).

Tromberg, B. J.

V. Venugopalan, J. S. You, and B. J. Tromberg, Phys. Rev. E 58, 2395 (1998).
[CrossRef]

Van Assendelft, O. W.

O. W. Van Assendelft, Spectrophotometry of Hemoglobin Derivatives (Charles C. Thomas, Springfield, Ill., 1970).

Venugopalan, V.

V. Venugopalan, J. S. You, and B. J. Tromberg, Phys. Rev. E 58, 2395 (1998).
[CrossRef]

Weinreb, A.

E. Loewinger, A. Gordon, A. Weinreb, and J. Gross, J. Appl. Physiol. 19, 1179 (1964).
[PubMed]

White, E. A.

Wray, S.

S. Wray, M. Cope, D. T. Delpy, J. S. Wyatt, and E. O. R. Reynolds, Biochim. Biophys. Acta 933, 184 (1988).
[CrossRef] [PubMed]

Wyatt, J. S.

S. Wray, M. Cope, D. T. Delpy, J. S. Wyatt, and E. O. R. Reynolds, Biochim. Biophys. Acta 933, 184 (1988).
[CrossRef] [PubMed]

You, J. S.

V. Venugopalan, J. S. You, and B. J. Tromberg, Phys. Rev. E 58, 2395 (1998).
[CrossRef]

Zijlstra, W. G.

W. G. Zijlstra, A. Bursma, and W. P. Meeuwsen-van der Roest, Clin. Chem. 37, 1633 (1991).
[PubMed]

Appl. Opt. (1)

Arch. Biochem. Biophys. (1)

P. Latimer and C. A. H. Eubanks, Arch. Biochem. Biophys. 98, 274 (1962).
[CrossRef] [PubMed]

Biochim. Biophys. Acta (2)

L. N. M. Duysens, Biochim. Biophys. Acta 19, 1 (1956).
[CrossRef] [PubMed]

S. Wray, M. Cope, D. T. Delpy, J. S. Wyatt, and E. O. R. Reynolds, Biochim. Biophys. Acta 933, 184 (1988).
[CrossRef] [PubMed]

Clin. Chem. (1)

W. G. Zijlstra, A. Bursma, and W. P. Meeuwsen-van der Roest, Clin. Chem. 37, 1633 (1991).
[PubMed]

IEEE Trans. Biomed. Eng. (1)

S. Takatani and M. D. Graham, IEEE Trans. Biomed. Eng. BME-25, 656 (1987).

J. Appl. Physiol. (1)

E. Loewinger, A. Gordon, A. Weinreb, and J. Gross, J. Appl. Physiol. 19, 1179 (1964).
[PubMed]

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

Methods Enzymol. (1)

G. N. Lapennas, J. M. Colacino, and J. Bonaventura, Methods Enzymol. 76, 449 (1981).
[CrossRef]

Microvasc. Res. (1)

S. Patel and R. N. Pittman, Microvasc. Res. 62, 63 (2001).
[CrossRef] [PubMed]

Phys. Med. Biol. (1)

E. L. Hull, M. G. Nichols, and T. H. Foster, Phys. Med. Biol. 43, 3381 (1998).
[CrossRef] [PubMed]

Phys. Rev. E (1)

V. Venugopalan, J. S. You, and B. J. Tromberg, Phys. Rev. E 58, 2395 (1998).
[CrossRef]

Other (3)

S. A. Prahl, “Optical properties of hemoglobin,” http://www.omlc.ogi.edu/spectra/hemoglobin/summary.html (retrieved July30, 2001).

E. M. Jope, in Hemoglobin, F. J. W. Roughton and J. C. Kendrew, eds. (Interscience, New York, 1949), pp. 205–219.

O. W. Van Assendelft, Spectrophotometry of Hemoglobin Derivatives (Charles C. Thomas, Springfield, Ill., 1970).

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Figures (3)

Fig. 1
Fig. 1

Value of Q resulting from the packaging of an absorber into a 1.6% suspension of 5µm-diameter spherical particles, plotted as a function of μa of the absorbing particles. Q is defined in Eq. (5).

Fig. 2
Fig. 2

Comparison of the spectra of 100µM hemoglobin in solution and suspended in spherical particles of 5µm diameter for (a) oxyhemoglobin and (b) deoxyhemoglobin.

Fig. 3
Fig. 3

Diffuse reflectance spectrum obtained from a partially deoxygenated erythrocyte phantom () and best fit of the P3-based fitting algorithm using basis spectra of hemoglobin in homogeneous solution () and corrected for the effects of pigment packaging ().

Equations (6)

Equations on this page are rendered with MathJax. Learn more.

T1=A-ap/A+ap/ATp=1-ap/A1-Tp.
T=T1NAl.
μasus=-NA ln1-ap/A1-TpNap1-Tp,
μasol=μaVParticlesVTotal=μaNAlvpAl=μaNvp,
Q=μasusμasol=1-Tpapvpμa.
Tp=22µar21-2µar+1exp-2μar,

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