June 2013
Volume 54, Issue 15
Free
ARVO Annual Meeting Abstract  |   June 2013
Characterization of lipids in human meibomian gland using coherent Raman scattering microscopy
Author Affiliations & Notes
  • Jeffrey Suhalim
    Beckman Laser Institute, University of California, Irvine, Irvine, CA
  • Yilu Xie
    Gavin Herbert Eye Institute, University of California, Irvine, Irvine, CA
  • Jeremiah Tao
    Gavin Herbert Eye Institute, University of California, Irvine, Irvine, CA
  • Donald Brown
    Gavin Herbert Eye Institute, University of California, Irvine, Irvine, CA
  • James Jester
    Gavin Herbert Eye Institute, University of California, Irvine, Irvine, CA
  • Eric Potma
    Beckman Laser Institute, University of California, Irvine, Irvine, CA
  • Footnotes
    Commercial Relationships Jeffrey Suhalim, None; Yilu Xie, None; Jeremiah Tao, None; Donald Brown, None; James Jester, None; Eric Potma, None
  • Footnotes
    Support None
Investigative Ophthalmology & Visual Science June 2013, Vol.54, 1486. doi:https://doi.org/
  • Views
  • Share
  • Tools
    • Alerts
      ×
      This feature is available to authenticated users only.
      Sign In or Create an Account ×
    • Get Citation

      Jeffrey Suhalim, Yilu Xie, Jeremiah Tao, Donald Brown, James Jester, Eric Potma; Characterization of lipids in human meibomian gland using coherent Raman scattering microscopy. Invest. Ophthalmol. Vis. Sci. 2013;54(15):1486. doi: https://doi.org/.

      Download citation file:


      © ARVO (1962-2015); The Authors (2016-present)

      ×
  • Supplements
Abstract

Purpose: While the association between meibomian gland dysfunction (MGD) and dry eye disease is well known, it is unclear how meibum lipid changes during the progression of the disease. Several studies have pointed out the importance of meibum composition in MGD; however, these studies primarily focused on studying lipids expressed from the meibomian gland. Hence, no information on compositional changes pertaining to the structure of the gland is available. In this study, we demonstrate the application of coherent Raman scattering (CRS) microscopy to characterize the compositional changes within the human meibomian gland.

Methods: Eyelid tissues from blepharoplasty patients were fixed in PBS with 2% PFA. Cryostat tissue sections were placed on a microscope slide, then imaged with a multimodal CRS microscope as described in [1]. The protein-to-lipid ratio (P/L) was obtained by tuning the laser beams to probe the vibrational signatures of protein and lipid. Specifically, the vibrations of methyl and amide-I were utilized to measure the contribution from protein, whereas the symmetric stretch and bending modes of methylene were used to probe lipid. In addition, the conjugated carbon-carbon vibration was monitored as a marker for carotenoid-like molecules. No labeling or staining was required in this study.

Results: In general, CRS analysis of the meibomian gland starting at the acinus and moving to the central duct showed a decreasing P/L ratio. Our preliminary results also showed differences in the P/L ratio between acini of the same gland. Concomitantly, Raman vibration of conjugated carbon-carbon bond was detected, hinting the presence of carotenoid-like molecules in the gland, as previously observed by Oshima et al. for expressed meibum [2].

Conclusions: CRS imaging enables compositional analysis in context of the native structure of the gland, complementing other studies. Our findings suggest that there is a maturational change in meibum with decreasing protein content as lipid moves toward the gland orifice and that there may be varying degrees of acinar activity within the gland. [1] Suhalim, J.L., et al., Biophysical Journal, 2012. 102(8): p. 1988-95. [2] Oshima, Y., et al., Current Eye Research, 2009. 34(10): p. 824-835.

Keywords: 551 imaging/image analysis: non-clinical • 486 cornea: tears/tear film/dry eye • 583 lipids  
×
×

This PDF is available to Subscribers Only

Sign in or purchase a subscription to access this content. ×

You must be signed into an individual account to use this feature.

×