Abstract

We present a method for selective detection of size-dependent scattering characteristics of epithelial cells in vivo based on polarized illumination and polarization sensitive detection of scattered light. We illustrate the method using phantoms designed to simulate squamous epithelial tissue and progressing to epithelial tissue in vitro and in vivo. Elastic light scattering spectroscopy with polarized illumination/detection dramatically reduces background signals due to both diffuse stromal scattering and hemoglobin absorption. Resulting spectra can be described as a linear combination of forward and backscattering components determined from Mie theory. Nuclear sizes and refractive indices extracted by fitting experimental spectra to this model agree well with previous measurements. Reflectance spectroscopy with polarized light can provide quantitative morphological information which could potentially be used for non-invasive detection of neoplastic changes.

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  1. American Cancer Society (1993) Cancer Facts and Figures. Publication No. 93-400M, No. 5008-03.
  2. J. R. Mourant, T. Fuselier, J. Boyer, T. M. Johnson, and I. J. Bigio, "Predictions and Measurements of Scattering and Absorption Over Broad Wavelength Ranges in Tissue Phantoms," Appl. Opt. 36, 949-57 (1997).
  3. L. T. Perelman, V. Backman, M. Wallace, G. Zonios, R. Manoharan, A. Nusrat, S. Shields, M. Seiler, C. Lima, T. Hamano, I. Itzkan, J. Van Dam, J. M. Crawford, and M. S. Feld, "Observation of Periodic Fine Structure in Reflectance from Biological Tissue: A New Technique for Measuring Nuclear Size Distribution," Phys. Rev. Lett. 80, 627-630 (1998).
  4. R. R. Anderson, "Polarized Light Examination and Photography of the Skin," Arch. Dermatol. 127, 1000-5 (1991).
  5. http://omlc.ogi.edu/news/feb98/polarization/index.html
  6. V. Backman, R. Gurjar, K. Badizadegan, I. Itzkan, R. Dasari, L. T. Perelman, and M. S. Feld, "Polarized Light Scattering Spectroscopy for Quantitative Measurements of Epithelial Cellular Structures In Situ," IEEE, J. Selected Topics in Quantum Electronics on Lasers in Medicine and Biology 5, (1999).
  7. T.M. Johnson and J.R. Mourant, "Polarized Wavelength-Dependant Measurements of Turbid Media," Opt. Express 4, 200-216 (1999). http://www.opticsexpress.org/oearchive/source/8492.htm
  8. V. Sankaran, K. Sch�nenberger, J. T. Walsh, Jr., and D. J. Maitland, "Polarization Discrimination of Coherently Propagating Light in Turbid Media," Appl. Opt. 38, 4252-4261 (1999).
  9. G. Jarry, E. Steimer, V. Damaschini, M. Epifanie, M. Jurczak, and R. Kaiser, "Coherence and Polarization of Light Propagating Through scattering Media and Biological Tissues," Appl. Opt. 37, 7357-7367 (1998).
  10. D. Bicout, C. Brosseau, A. S. Martinez, J. M. Schmitt, "Depolarization of Multiply Scattered Waves by Spherical Diffusers: Influence of the Size Parameter," Phys. Rev. E 49, 1767-1770 (1994).
  11. R. Barer, "Refractometry and Interferometry of Living Cells," J. Opt. Soc. Am. 47, 545-56 (1957).
  12. C. Smithpeter, A. Dunn, R. Drezek, T. Collier, and R. Richards-Kortum, "Near Real Time Confocal Microscopy of Cultured Amelanotic Cells: Sources of Signal, Contrast Agents and Limits of Contrast," J. Biomed. Opt. 3, 429-36 (1998).
  13. H.D. Young. Statistical Treatment of Experimental Data. (McGraw-Hill, New York, NY, 1962).
  14. H. van de Hulst. Light Scattering by Small Particles. (Dover, New York, NY, 1957).
  15. R. Kudo, S. Sagae, O. Hayakawa, E. Ito, E. Horimoto, and M. Hashimoto, "Morphology of Adenocarcinoma in Situ and Microinvasive Adenocarcinoma of the Uterine Cervix. A Cytologic and Ultrastructural Study," Acta Cytol. 35, 109-16 (1991).
  16. N. Wheeler, S. C. Suffin, T. L. Hall, and D. L. Rosenthal "Predictions of Cervical Neoplasia Diagnosis Groups. Discriminant Analysis on Digitized Cell Images," Analyt. Quant. Cytol. Histol. 9, 169-81 (1987).
  17. E. Artacho-Perula, R. Roldan-Villalobos, J. Salas-Molina, and R. Vaamonde-Lemos, "Histomorphometry of Normal and Abnormal Cervical Samples," Analyt. Quant. Cytol. Histol. 15, 290-97 (1993).
  18. L. Burke, D. A. Antonioli, B. S. Ducatman, Colposcopy. Text and Atlas, (Appleton & Lance, CA 1991).
  19. A. Dunn, C. Smithpeter, A. J. Welch, R. Richards-Kortum, "FDTD Simulation of Light Scattering from Single Cells," J. Biomed. Optic. 2, 262-66 (1997).
  20. R. Drezek, A. Dunn, and R. Richards-Kortum, "Light Scattering from Cells: Finite-Difference Time-Domain Simulations and Goniometric Measurements," Appl. Opt. 38, 3651-3661 (1999).
  21. J. R. Mourant, J. P. freyer, A. H. Hielscher, A. A. Eick, D. Shen, and T. M. Johnson, "Mechanism of Light Scattering from Biological Cells Relevant to Noninvasive Optical-Tissue Diagnostics," Appl. Opt. 37, 3586-3593 (1998).
  22. A. Brunsting and P. F. Mullaney, "Differential Light Scattering from Spherical Mammalian Cells," Biophys. J. 14, 439-453 (1974).
  23. J. M. Schmitt and G. Kumar, "Optical Scattering Properties of Soft Tissue: a Discrete Particle Model," Appl. Opt. 37, 2788-2797 (1998).
  24. S. Asano and M. Sato, "Light Scattering by Randomly Oriented Spheroidal Perticles," Appl. Opt. 19, 962- 974 (1980).

Other

American Cancer Society (1993) Cancer Facts and Figures. Publication No. 93-400M, No. 5008-03.

J. R. Mourant, T. Fuselier, J. Boyer, T. M. Johnson, and I. J. Bigio, "Predictions and Measurements of Scattering and Absorption Over Broad Wavelength Ranges in Tissue Phantoms," Appl. Opt. 36, 949-57 (1997).

L. T. Perelman, V. Backman, M. Wallace, G. Zonios, R. Manoharan, A. Nusrat, S. Shields, M. Seiler, C. Lima, T. Hamano, I. Itzkan, J. Van Dam, J. M. Crawford, and M. S. Feld, "Observation of Periodic Fine Structure in Reflectance from Biological Tissue: A New Technique for Measuring Nuclear Size Distribution," Phys. Rev. Lett. 80, 627-630 (1998).

R. R. Anderson, "Polarized Light Examination and Photography of the Skin," Arch. Dermatol. 127, 1000-5 (1991).

http://omlc.ogi.edu/news/feb98/polarization/index.html

V. Backman, R. Gurjar, K. Badizadegan, I. Itzkan, R. Dasari, L. T. Perelman, and M. S. Feld, "Polarized Light Scattering Spectroscopy for Quantitative Measurements of Epithelial Cellular Structures In Situ," IEEE, J. Selected Topics in Quantum Electronics on Lasers in Medicine and Biology 5, (1999).

T.M. Johnson and J.R. Mourant, "Polarized Wavelength-Dependant Measurements of Turbid Media," Opt. Express 4, 200-216 (1999). http://www.opticsexpress.org/oearchive/source/8492.htm

V. Sankaran, K. Sch�nenberger, J. T. Walsh, Jr., and D. J. Maitland, "Polarization Discrimination of Coherently Propagating Light in Turbid Media," Appl. Opt. 38, 4252-4261 (1999).

G. Jarry, E. Steimer, V. Damaschini, M. Epifanie, M. Jurczak, and R. Kaiser, "Coherence and Polarization of Light Propagating Through scattering Media and Biological Tissues," Appl. Opt. 37, 7357-7367 (1998).

D. Bicout, C. Brosseau, A. S. Martinez, J. M. Schmitt, "Depolarization of Multiply Scattered Waves by Spherical Diffusers: Influence of the Size Parameter," Phys. Rev. E 49, 1767-1770 (1994).

R. Barer, "Refractometry and Interferometry of Living Cells," J. Opt. Soc. Am. 47, 545-56 (1957).

C. Smithpeter, A. Dunn, R. Drezek, T. Collier, and R. Richards-Kortum, "Near Real Time Confocal Microscopy of Cultured Amelanotic Cells: Sources of Signal, Contrast Agents and Limits of Contrast," J. Biomed. Opt. 3, 429-36 (1998).

H.D. Young. Statistical Treatment of Experimental Data. (McGraw-Hill, New York, NY, 1962).

H. van de Hulst. Light Scattering by Small Particles. (Dover, New York, NY, 1957).

R. Kudo, S. Sagae, O. Hayakawa, E. Ito, E. Horimoto, and M. Hashimoto, "Morphology of Adenocarcinoma in Situ and Microinvasive Adenocarcinoma of the Uterine Cervix. A Cytologic and Ultrastructural Study," Acta Cytol. 35, 109-16 (1991).

N. Wheeler, S. C. Suffin, T. L. Hall, and D. L. Rosenthal "Predictions of Cervical Neoplasia Diagnosis Groups. Discriminant Analysis on Digitized Cell Images," Analyt. Quant. Cytol. Histol. 9, 169-81 (1987).

E. Artacho-Perula, R. Roldan-Villalobos, J. Salas-Molina, and R. Vaamonde-Lemos, "Histomorphometry of Normal and Abnormal Cervical Samples," Analyt. Quant. Cytol. Histol. 15, 290-97 (1993).

L. Burke, D. A. Antonioli, B. S. Ducatman, Colposcopy. Text and Atlas, (Appleton & Lance, CA 1991).

A. Dunn, C. Smithpeter, A. J. Welch, R. Richards-Kortum, "FDTD Simulation of Light Scattering from Single Cells," J. Biomed. Optic. 2, 262-66 (1997).

R. Drezek, A. Dunn, and R. Richards-Kortum, "Light Scattering from Cells: Finite-Difference Time-Domain Simulations and Goniometric Measurements," Appl. Opt. 38, 3651-3661 (1999).

J. R. Mourant, J. P. freyer, A. H. Hielscher, A. A. Eick, D. Shen, and T. M. Johnson, "Mechanism of Light Scattering from Biological Cells Relevant to Noninvasive Optical-Tissue Diagnostics," Appl. Opt. 37, 3586-3593 (1998).

A. Brunsting and P. F. Mullaney, "Differential Light Scattering from Spherical Mammalian Cells," Biophys. J. 14, 439-453 (1974).

J. M. Schmitt and G. Kumar, "Optical Scattering Properties of Soft Tissue: a Discrete Particle Model," Appl. Opt. 37, 2788-2797 (1998).

S. Asano and M. Sato, "Light Scattering by Randomly Oriented Spheroidal Perticles," Appl. Opt. 19, 962- 974 (1980).

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