June 2013
Volume 54, Issue 15
Free
ARVO Annual Meeting Abstract  |   June 2013
Imaging the Effects of Prostaglandins on Cultured Trabecular Meshwork Cells by Coherent Anti-Stokes Raman Scattering (CARS)
Author Affiliations & Notes
  • David Ammar
    Ophthalmology, Univ of Colorado Denver School of Medicine, Aurora, CO
  • Omid Masihzadeh
    Ophthalmology, Univ of Colorado Denver School of Medicine, Aurora, CO
  • Malik Kahook
    Ophthalmology, Univ of Colorado Denver School of Medicine, Aurora, CO
  • Tim Lei
    Electrical Engineering, Univ of Colorado Denver, Denver, CO
  • Footnotes
    Commercial Relationships David Ammar, U.S. Patent Application #13/700,682 (P); Omid Masihzadeh, U.S. Patent Application #13/700,682 (P); Malik Kahook, Alcon (C), Allergan (C), Merck (C), B&L (C), Glaukos (C), Ivantis (C), ClarVista Medical (P), Dose Medical (P), AMO (P), Genentech (F), Regeneron (F); Tim Lei, U.S. Patent Application #13/700,682 (P)
  • Footnotes
    Support None
Investigative Ophthalmology & Visual Science June 2013, Vol.54, 3530. doi:https://doi.org/
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    • Get Citation

      David Ammar, Omid Masihzadeh, Malik Kahook, Tim Lei; Imaging the Effects of Prostaglandins on Cultured Trabecular Meshwork Cells by Coherent Anti-Stokes Raman Scattering (CARS). Invest. Ophthalmol. Vis. Sci. 2013;54(15):3530. doi: https://doi.org/.

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

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Abstract

Purpose: To non-destructively monitor morphological changes to the lipid membranes of living primary cultures of human trabecular meshwork cells (hTMC) without the application of exogenous label.

Methods: hTMC were imaged using a non-linear optical technique, coherent anti-Stokes Raman scattering (CARS), after treatment with a commercial formulation of latanoprost. Untreated cells and cells treated with vehicle containing the preservative benzalkonium chloride (BAK) were imaged as controls. Cells were fixed, stained with the fluorescent lipid dye Nile Red and imaged by conventional confocal microscopy to verify lipid membrane structures.

Results: Analysis of CARS images of hTMC treated for 24 hours with 0.5 µg/ml latanoprost revealed multiple intracellular lipid membranes absent from untreated or BAK-treated hTMC. Treatment of hTMC with sodium fluoride or ouabain, agents shown to cause morphological changes to hTMC, also did not induce formation of intracellular lipid membrane.

Conclusions: CARS microscopy detected changes in living hTMC morphology that was validated by subsequent histological stain. Prostaglandin-induced changes to hTMC involved rearrangement of lipid membrane within these cells. These in vitro results identify a novel biological response to a class of anti-glaucoma drugs, and further experiments are needed to establish how this effect is involved in the hypotensive action of prostaglandins in vivo.

Keywords: 735 trabecular meshwork • 568 intraocular pressure • 596 microscopy: confocal/tunneling  
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