In in vivo spectroscopy, there are differences between individual subjects in parameters such as tissue scattering and sample concentration. We propose a method that can provide the absolute value of a particular substance concentration, independent of these individual differences. Thus, it is not necessary to use the typical statistical calibration curve, which assumes an average level of scattering and an averaged concentration over individual subjects. This method is expected to greatly reduce the difficulties encountered during in vivo measurements. As an example, for in vivo absorption spectroscopy, the method was applied to the reflectance measurement in retinal vessels to monitor their oxygen saturation levels. This method was then validated by applying it to the tissue phantom under a variety of absorbance values and scattering efficiencies.

© 2012 OSA

1. M. Born and E. Wolf, “Beam propagation in an absorbing medium,” in Principles of Optics 7th Ed. (Cambridge University Press, Cambridge, 2002).
2. J. H. Lambert, Photometria sive de mensura et gradibus luminus, colorum et umbrae (1760) [Published in German by E. Anding under the title Lambert’s Photometrie, (Verlag von Wilhelm Engelmann, Leipzig, 1892)].
3. A. Beer, “Bestimmung der Absorption des rothen Lichts in farbigen Flüssigkeiten,” Annal. Phys. Chem.86, 78–88 (1852).
4. J. M. Beach, K. J. Schwenzer, S. Srinivas, D. Kim, and J. S. Tiedeman, “Oximetry of retinal vessels by dual-wavelength imaging: calibration and influence of pigmentation,” J. Appl. Physiol.86(2), 748–758 (1999). [PubMed]
5. M. H. Smith, K. R. Denninghoff, A. Lompado, and L. W. Hillman, “Effect of multiple light paths on retinal vessel oximetry,” Appl. Opt.39(7), 1183–1193 (2000). [CrossRef] [PubMed]
6. A. Kienle, M. S. Patterson, N. Dögnitz, R. Bays, G. Wagnieres, and H. van den Bergh, “Noninvasive determination of the optical properties of two-layered turbid media,” Appl. Opt.37(4), 779–791 (1998). [CrossRef] [PubMed]
7. H. M. Heise, “Applications of near-infrared spectroscopy in medical sciences,” Near-Infrared Spectroscopy, H. W. Siesler, Y. Ozeki, S. Kawata, and H. M. Heise ed. (Wiley VCH Verlag GmbH, Weinheim, 2002).
8. A. J. Cohen and R. A. Laing, “Multiple scattering analysis of retinal blood oximetry,” IEEE Trans. Biomed. Eng.BME-23(5), 391–400 (1976). [CrossRef] [PubMed]
9. S. Prahl, “Optical absorption of hemoglobin.” http://omlc.ogi.edu/spectra/hemoglobin .
10. P. Kubelka and F. Munk, ““Ein Beitrag zur Optik der Farbanstriche,” Zeits. f,” Tech. Phys.12, 593–601 (1931).
11. S. Traustason, A. S. Jensen, H. S. Arvidsson, I. C. Munch, L. Søndergaard, and M. Larsen, “Retinal oxygen saturation in patients with systemic hypoxemia,” Invest. Ophthalmol. Vis. Sci.52(8), 5064–5067 (2011). [CrossRef] [PubMed]
12. Medical electrical equipment Part 2: Particular requirements for the safety of diagnostic and therapeutic laser equipment, IEC 60101–2-22 and IEC 60825–1. Japanese Industrial Standards, JIS C 6802–2005.
13. J. V. B. Soares, J. J. G. Leandro, R. M. Cesar, H. F. Jelinek, and M. J. Cree, “Retinal vessel segmentation using the 2-D Gabor wavelet and supervised classification,” IEEE Trans. Med. Imaging25(9), 1214–1222 (2006). [CrossRef] [PubMed]
14. F. C. Mokken, F. J. M. van der Waart, C. P. Henny, P. T. Goedhart, and A. W. Gelb, “Differences in peripheral arterial and venous hemorheologic parameters,” Ann. Hematol.73(3), 135–137 (1996). [CrossRef] [PubMed]
15. J. M. Steinke and A. P. Shepherd, “Comparison of Mie theory and the light scattering of red blood cells,” Appl. Opt.27(19), 4027–4033 (1988). [CrossRef] [PubMed]
16. M. S. Patterson, B. C. Wilson, and D. R. Wyman, “The propagation of optical radiation in tissue I. Models of radiation transport and their application,” Lasers Med. Sci.6(2), 155–168 (1991). [CrossRef]
17. D. Link, C. Strohmaier, B. U. Seifert, T. Riemer, H. A. Reitsamer, J. Haueisen, and W. Vilser, “Novel non-contact retina camera for the rat and its application to dynamic retinal vessel analysis,” Biomed. Opt. Express2(11), 3094–3108 (2011). [CrossRef] [PubMed]
18. J. W. Severinghaus and Y. Honda, “History of blood gas analysis. VII. Pulse oximetry,” J. Clin. Monit.3(2), 135–138 (1987). [CrossRef] [PubMed]

Arvidsson, H. S.
Bays, R.
Beach, J. M.
Beer, A.
Cesar, R. M.
Cohen, A. J.
Cree, M. J.
Denninghoff, K. R.
Dögnitz, N.
Gelb, A. W.
Goedhart, P. T.
Haueisen, J.
Henny, C. P.
Hillman, L. W.
Honda, Y.
Jelinek, H. F.
Jensen, A. S.
Kienle, A.
Kim, D.
Kubelka, P.
Laing, R. A.
Larsen, M.
Leandro, J. J. G.
Link, D.
Lompado, A.
Mokken, F. C.
Munch, I. C.
Munk, F.
Patterson, M. S.
Reitsamer, H. A.
Riemer, T.
Schwenzer, K. J.
Seifert, B. U.
Severinghaus, J. W.
Shepherd, A. P.
Smith, M. H.
Soares, J. V. B.
Søndergaard, L.
Srinivas, S.
Steinke, J. M.
Strohmaier, C.
Tiedeman, J. S.
Traustason, S.
van den Bergh, H.
van der Waart, F. J. M.
Vilser, W.
Wagnieres, G.
Wilson, B. C.
Wyman, D. R.

Ann. Hematol.

• F. C. Mokken, F. J. M. van der Waart, C. P. Henny, P. T. Goedhart, and A. W. Gelb, “Differences in peripheral arterial and venous hemorheologic parameters,” Ann. Hematol.73(3), 135–137 (1996). [CrossRef] [PubMed]

Annal. Phys. Chem.

• A. Beer, “Bestimmung der Absorption des rothen Lichts in farbigen Flüssigkeiten,” Annal. Phys. Chem.86, 78–88 (1852).

Appl. Opt.

• M. H. Smith, K. R. Denninghoff, A. Lompado, and L. W. Hillman, “Effect of multiple light paths on retinal vessel oximetry,” Appl. Opt.39(7), 1183–1193 (2000). [CrossRef] [PubMed]
• A. Kienle, M. S. Patterson, N. Dögnitz, R. Bays, G. Wagnieres, and H. van den Bergh, “Noninvasive determination of the optical properties of two-layered turbid media,” Appl. Opt.37(4), 779–791 (1998). [CrossRef] [PubMed]
• J. M. Steinke and A. P. Shepherd, “Comparison of Mie theory and the light scattering of red blood cells,” Appl. Opt.27(19), 4027–4033 (1988). [CrossRef] [PubMed]

Biomed. Opt. Express

• D. Link, C. Strohmaier, B. U. Seifert, T. Riemer, H. A. Reitsamer, J. Haueisen, and W. Vilser, “Novel non-contact retina camera for the rat and its application to dynamic retinal vessel analysis,” Biomed. Opt. Express2(11), 3094–3108 (2011). [CrossRef] [PubMed]

IEEE Trans. Biomed. Eng.

• A. J. Cohen and R. A. Laing, “Multiple scattering analysis of retinal blood oximetry,” IEEE Trans. Biomed. Eng.BME-23(5), 391–400 (1976). [CrossRef] [PubMed]

IEEE Trans. Med. Imaging

• J. V. B. Soares, J. J. G. Leandro, R. M. Cesar, H. F. Jelinek, and M. J. Cree, “Retinal vessel segmentation using the 2-D Gabor wavelet and supervised classification,” IEEE Trans. Med. Imaging25(9), 1214–1222 (2006). [CrossRef] [PubMed]

Invest. Ophthalmol. Vis. Sci.

• S. Traustason, A. S. Jensen, H. S. Arvidsson, I. C. Munch, L. Søndergaard, and M. Larsen, “Retinal oxygen saturation in patients with systemic hypoxemia,” Invest. Ophthalmol. Vis. Sci.52(8), 5064–5067 (2011). [CrossRef] [PubMed]

J. Appl. Physiol.

• J. M. Beach, K. J. Schwenzer, S. Srinivas, D. Kim, and J. S. Tiedeman, “Oximetry of retinal vessels by dual-wavelength imaging: calibration and influence of pigmentation,” J. Appl. Physiol.86(2), 748–758 (1999). [PubMed]

J. Clin. Monit.

• J. W. Severinghaus and Y. Honda, “History of blood gas analysis. VII. Pulse oximetry,” J. Clin. Monit.3(2), 135–138 (1987). [CrossRef] [PubMed]

Lasers Med. Sci.

• M. S. Patterson, B. C. Wilson, and D. R. Wyman, “The propagation of optical radiation in tissue I. Models of radiation transport and their application,” Lasers Med. Sci.6(2), 155–168 (1991). [CrossRef]

Tech. Phys.

• P. Kubelka and F. Munk, ““Ein Beitrag zur Optik der Farbanstriche,” Zeits. f,” Tech. Phys.12, 593–601 (1931).

Other

• Medical electrical equipment Part 2: Particular requirements for the safety of diagnostic and therapeutic laser equipment, IEC 60101–2-22 and IEC 60825–1. Japanese Industrial Standards, JIS C 6802–2005.
• M. Born and E. Wolf, “Beam propagation in an absorbing medium,” in Principles of Optics 7th Ed. (Cambridge University Press, Cambridge, 2002).
• J. H. Lambert, Photometria sive de mensura et gradibus luminus, colorum et umbrae (1760) [Published in German by E. Anding under the title Lambert’s Photometrie, (Verlag von Wilhelm Engelmann, Leipzig, 1892)].
• S. Prahl, “Optical absorption of hemoglobin.” http://omlc.ogi.edu/spectra/hemoglobin .
• H. M. Heise, “Applications of near-infrared spectroscopy in medical sciences,” Near-Infrared Spectroscopy, H. W. Siesler, Y. Ozeki, S. Kawata, and H. M. Heise ed. (Wiley VCH Verlag GmbH, Weinheim, 2002).

2011, Traustason, Invest. Ophthalmol. Vis. Sci.
2011, Link, Biomed. Opt. Express
2006, Soares, IEEE Trans. Med. Imaging
2000, Smith, Appl. Opt.
1999, Beach, J. Appl. Physiol.
1998, Kienle, Appl. Opt.
1996, Mokken, Ann. Hematol.
1991, Patterson, Lasers Med. Sci.
1988, Steinke, Appl. Opt.
1987, Severinghaus, J. Clin. Monit.
1976, Cohen, IEEE Trans. Biomed. Eng.
1931, Kubelka, Tech. Phys.
1852, Beer, Annal. Phys. Chem.

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