Determination of epithelial tissue scattering coefficient using confocal microscopy

Most models of light propagation through tissue assume the scattering properties of the various tissue layers are the same. The authors present evidence that the scattering coefficient of normal cervical epithelium is significantly lower than values previously reported for bulk epithelial tissue. They estimated the scattering coefficient of normal and precancerous cervical epithelium using measurements of the reflectance as a function of depth from confocal images. Reflectance measurements were taken from ex vivo cervical biopsies and fit to an exponential function based upon Beer's law attenuation. The mean scattering coefficients derived were 22 cm/sup -1/ for normal tissue and 69 cm/sup -1/ for precancerous tissue. These values are significantly lower than previously reported for bulk epithelial tissues and suggest that scattering of bulk tissue is dominated by the stroma. They also suggest that computational models to describe light propagation in epithelial tissue must incorporate different scattering coefficients for the epithelium and stroma. Further, the lower scattering of the epithelium suggests greater probing depths for fiber optic probes used by optical diagnostic devices which measure reflectance and fluorescence in epithelial tissue. The difference in scattering between normal and precancerous tissue is attributed to increased nuclear size, optical density, and chromatin texture. The scattering coefficients measured here are consistent with predictions of numerical solutions to Maxwell's equations for epithelial cell scattering.

[1]  R. Richards-Kortum,et al.  Light scattering from cervical cells throughout neoplastic progression: influence of nuclear morphology, DNA content, and chromatin texture. , 2003, Journal of biomedical optics.

[2]  R. Richards-Kortum,et al.  Microanatomical and Biochemical Origins of Normal and Precancerous Cervical Autofluorescence Using Laser-scanning Fluorescence Confocal Microscopy¶ , 2003, Photochemistry and photobiology.

[3]  B W Pogue,et al.  Fiber-optic bundle design for quantitative fluorescence measurement from tissue. , 1998, Applied optics.

[4]  M S Patterson,et al.  Optical properties of normal and diseased human breast tissues in the visible and near infrared. , 1990, Physics in medicine and biology.

[5]  T Joshua Pfefer,et al.  Multiple-fiber probe design for fluorescence spectroscopy in tissue. , 2002, Applied optics.

[6]  N Kollias,et al.  Spectroscopic measurement of diffuse reflectance for enhanced detection of bladder carcinoma. , 1998, Urology.

[7]  R. Richards-Kortum,et al.  Light scattering from normal and dysplastic cervical cells at different epithelial depths: finite-difference time-domain modeling with a perfectly matched layer boundary condition. , 2003, Journal of biomedical optics.

[8]  B. Tromberg,et al.  Imaging cells and extracellular matrix in vivo by using second-harmonic generation and two-photon excited fluorescence , 2002, Proceedings of the National Academy of Sciences of the United States of America.

[9]  R. Lotan,et al.  Autofluorescence Microscopy of Fresh Cervical-Tissue Sections Reveals Alterations in Tissue Biochemistry with Dysplasia¶ , 2001, Photochemistry and photobiology.

[10]  J D Pitts,et al.  Autofluorescence characteristics of immortalized and carcinogen-transformed human bronchial epithelial cells. , 2001, Journal of biomedical optics.

[11]  R Richards-Kortum,et al.  Reflectance spectroscopy with polarized light: is it sensitive to cellular and nuclear morphology. , 1999, Optics express.

[12]  Wolfgang Rudolph,et al.  Comparative study of confocal and heterodyne microscopy for imaging through scattering media , 1996 .

[13]  Optical Tomography and Spectroscopy of Tissue: Theory, Instrumentation, Model, and Human Studies II , 1997 .

[14]  M. Kohl,et al.  Near-infrared optical properties of ex vivo human skin and subcutaneous tissues measured using the Monte Carlo inversion technique. , 1998, Physics in medicine and biology.

[15]  H Key,et al.  Optical attenuation characteristics of breast tissues at visible and near-infrared wavelengths. , 1991, Physics in medicine and biology.

[16]  Irene Georgakoudi,et al.  Trimodal spectroscopy for the detection and characterization of cervical precancers in vivo. , 2002, American journal of obstetrics and gynecology.

[17]  N. Nishioka,et al.  Identification of Colonic Dysplasia and Neoplasia by Diffuse Reflectance Spectroscopy and Pattern Recognition Techniques , 1998 .

[18]  J. Mourant,et al.  Ultraviolet and visible spectroscopies for tissue diagnostics: fluorescence spectroscopy and elastic-scattering spectroscopy. , 1997, Physics in medicine and biology.

[19]  S. Thennadil,et al.  Optical properties of human skin in the near infrared wavelength range of 1000 to 2200 nm. , 2001, Journal of biomedical optics.

[20]  M. Sporn,et al.  Recent advances in chemoprevention of cancer. , 1997, Science.

[21]  J. Pickering,et al.  Double-integrating-sphere system for measuring the optical properties of tissue. , 1993, Applied optics.

[22]  A. Welch,et al.  Determining the optical properties of turbid mediaby using the adding-doubling method. , 1993, Applied optics.

[23]  D I McLean,et al.  Rapid near-infrared Raman spectroscopy system for real-time in vivo skin measurements. , 2001, Optics letters.

[24]  J. Schmitt,et al.  Confocal microscopy in turbid media. , 1994, Journal of the Optical Society of America. A, Optics, image science, and vision.

[25]  E. Sevick-Muraca,et al.  Quantitative optical spectroscopy for tissue diagnosis. , 1996, Annual review of physical chemistry.

[26]  M. Rajadhyaksha,et al.  Confocal examination of nonmelanoma cancers in thick skin excisions to potentially guide mohs micrographic surgery without frozen histopathology. , 2001, The Journal of investigative dermatology.

[27]  G. Zonios,et al.  Diffuse reflectance spectroscopy of human adenomatous colon polyps in vivo. , 1999, Applied optics.

[28]  Design and performance of an endoscopic OCT system for in vivo studies of human mucosa , 1998, Technical Digest. Summaries of Papers Presented at the Conference on Lasers and Electro-Optics. Conference Edition. 1998 Technical Digest Series, Vol.6 (IEEE Cat. No.98CH36178).

[29]  Robert A. Glasgold,et al.  Tissue autofluorescence as an intermediate endpoint in NMBA-induced esophageal carcinogenesis. , 1994, Cancer letters.

[30]  P. Monnier,et al.  Clinical determination of tissue optical properties by endoscopic spatially resolved reflectometry. , 1996, Applied optics.

[31]  A. Halpern,et al.  Detection of clinically amelanotic malignant melanoma and assessment of its margins by in vivo confocal scanning laser microscopy. , 2001, Archives of dermatology.

[32]  R. Alfano,et al.  Optical spectroscopic diagnosis of cancer and normal breast tissues , 1989 .

[33]  Departement Frauenheilkunde,et al.  Quantitative near-infrared spectroscopy of cervical dysplasia in vivo , 1999 .

[34]  B Palcic,et al.  Optical properties of normal and carcinomatous bronchial tissue. , 1994, Applied optics.

[35]  A. Lacy,et al.  Near real time confocal microscopy of amelanotic tissue: detection of dysplasia in ex-vivo cervical tissue , 2002, Proceedings of the Second Joint 24th Annual Conference and the Annual Fall Meeting of the Biomedical Engineering Society] [Engineering in Medicine and Biology.

[36]  K. Badizadegan,et al.  Fluorescence, reflectance, and light-scattering spectroscopy for evaluating dysplasia in patients with Barrett's esophagus. , 2001, Gastroenterology.

[37]  R. Richards-Kortum,et al.  Raman spectroscopy for the detection of cancers and precancers. , 1996, Journal of biomedical optics.

[38]  R. Hunter Hemoglobin oxygenation of a two-layer tissue-simulating phantom from time-resolved reflectance , 2001 .

[39]  J. Fujimoto,et al.  Optical coherence tomography for optical biopsy. Properties and demonstration of vascular pathology. , 1996, Circulation.

[40]  I J Bigio,et al.  Spectroscopic diagnosis of bladder cancer with elastic light scattering , 1995, Lasers in surgery and medicine.

[41]  J. Mourant,et al.  Predictions and measurements of scattering and absorption over broad wavelength ranges in tissue phantoms. , 1997, Applied optics.

[42]  S L Jacques,et al.  Measurement of tissue optical properties by the use of oblique-incidence optical fiber reflectometry. , 1997, Applied optics.

[43]  Nirmala Ramanujam,et al.  Relationship between depth of a target in a turbid medium and fluorescence measured by a variable-aperture method. , 2002, Optics letters.

[44]  R. Richards-Kortum,et al.  Penetration depth limits of in vivo confocal reflectance imaging , 1997, CLEO '97., Summaries of Papers Presented at the Conference on Lasers and Electro-Optics.

[45]  A. Welch,et al.  A review of the optical properties of biological tissues , 1990 .

[46]  K. Schomacker,et al.  Laser induced autofluorescence diagnosis of bladder cancer. , 1996, The Journal of urology.