Picosecond spectral coherent anti-Stokes Raman scattering imaging with principal component analysis of meibomian glands

• Published in: Journal of Biomedical Optics Vol. 16; no. 2; p. 021104
• Main Authors: Lin, Chia-Yu, Potma, Eric O, Suhalim, Jeffrey L, Nien, Chyong Ly, Jester, James V, Miljkovic, Milo D, Diem, Max
• Format: Journal Article
• Language: English
• Published: United States Society of Photo-Optical Instrumentation Engineers (SPIE) 01-02-2011
• Subjects: Animals; Cars; Ducts; Image Enhancement - methods; Image Interpretation, Computer-Assisted - methods; Imaging; Lipids; Meibomian Glands - cytology; Mice; Microscopy - methods; Principal Component Analysis; Raman scattering; Reproducibility of Results; Sensitivity and Specificity; Special Section on Coherent Raman Imaging Techniques and Biomedical Applications; Spectra; Spectrum Analysis, Raman - methods; Tomography, Optical Coherence - methods; Utilities
• ISSN: 1083-3668; 1560-2281
• DOI: 10.1117/1.3533716

The lipid distribution in the mouse meibomian gland was examined with picosecond spectral anti-Stokes Raman scattering (CARS) imaging. Spectral CARS data sets were generated by imaging specific localized regions of the gland within tissue sections at consecutive Raman shifts in the CH
stretching vibrational range. Spectral differences between the location specific CARS spectra obtained in the lipid-rich regions of the acinus and the central duct were observed, which were confirmed with a Raman microspectroscopic analysis, and attributed to meibum lipid modifications within the gland. A principal component analysis of the spectral data set reveals changes in the CARS spectrum when transitioning from the acini to the central duct. These results demonstrate the utility of picosecond spectral CARS imaging combined with multivariate analysis for assessing differences in the distribution and composition of lipids in tissues.