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Journal of Biomedical Optics • Open Access

Quantitative polarized Raman spectroscopy in highly turbid bone tissue
Author(s): Mekhala Raghavan; Nadder D. Sahar; Robert H. Wilson; Mary-Ann Mycek; Nancy Pleshko; David H. Kohn; Michael D. Morris

Paper Abstract

Polarized Raman spectroscopy allows measurement of molecular orientation and composition and is widely used in the study of polymer systems. Here, we extend the technique to the extraction of quantitative orientation information from bone tissue, which is optically thick and highly turbid. We discuss multiple scattering effects in tissue and show that repeated measurements using a series of objectives of differing numerical apertures can be employed to assess the contributions of sample turbidity and depth of field on polarized Raman measurements. A high numerical aperture objective minimizes the systematic errors introduced by multiple scattering. We test and validate the use of polarized Raman spectroscopy using wild-type and genetically modified (oim/oim model of osteogenesis imperfecta) murine bones. Mineral orientation distribution functions show that mineral crystallites are not as well aligned (p<0.05) in oim/oim bones (28±3 deg) compared to wild-type bones (22±3 deg), in agreement with small-angle X-ray scattering results. In wild-type mice, backbone carbonyl orientation is 76±2 deg and in oim/oim mice, it is 72±4 deg (p>0.05). We provide evidence that simultaneous quantitative measurements of mineral and collagen orientations on intact bone specimens are possible using polarized Raman spectroscopy.

Paper Details

Date Published: 1 May 2010
PDF: 7 pages
J. Biomed. Opt. 15(3) 037001 doi: 10.1117/1.3426310
Published in: Journal of Biomedical Optics Volume 15, Issue 3
Show Author Affiliations
Mekhala Raghavan, Univ. of Michigan (United States)
Nadder D. Sahar
Robert H. Wilson, Univ. of Michigan (United States)
Mary-Ann Mycek, Univ. of Michigan (United States)
Nancy Pleshko, Temple Univ. (United States)
David H. Kohn, Univ. of Michigan (United States)
Michael D. Morris, Univ. of Michigan (United States)

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