May 2005
Volume 46, Issue 13
Free
ARVO Annual Meeting Abstract  |   May 2005
A Systematic, Experimental Method of Screening Novel Retinal Vital Stains
Author Affiliations & Notes
  • T.L. Jackson
    Ophthalmology, Rayne Institute, London, United Kingdom
    Moorfields Eye Hospital, London, United Kingdom
  • L. Griffin
    Imaging Sciences, King's College London, London, United Kingdom
  • B. Vote
    Ophthalmology, Rayne Institute, London, United Kingdom
    Moorfields Eye Hospital, London, United Kingdom
  • J. Hillenkamp
    Ophthalmology, Rayne Institute, London, United Kingdom
    Eye Hospital, University of Regensburg, Regensburg, Germany
  • J. Marshall
    Ophthalmology, Rayne Institute, London, United Kingdom
  • Footnotes
    Commercial Relationships  T.L. Jackson, None; L. Griffin, None; B. Vote, None; J. Hillenkamp, None; J. Marshall, None.
  • Footnotes
    Support  Special Trustees of Guys and St Thomas' Hospital, Allerton Trust, DFG grant Hi 758/1–1
Investigative Ophthalmology & Visual Science May 2005, Vol.46, 5530. doi:
  • Views
  • Share
  • Tools
    • Alerts
      ×
      This feature is available to authenticated users only.
      Sign In or Create an Account ×
    • Get Citation

      T.L. Jackson, L. Griffin, B. Vote, J. Hillenkamp, J. Marshall; A Systematic, Experimental Method of Screening Novel Retinal Vital Stains . Invest. Ophthalmol. Vis. Sci. 2005;46(13):5530.

      Download citation file:


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

      ×
  • Supplements
Abstract

Abstract: : Purpose: To develop a method of testing the potential surgical utility of novel retinal vital stains. Methods: Bovine retina was exposed to test agents at a range of concentrations. Masked observers determined the minimum dye concentration that reliably stained the retina, defined as the minimum visible concentration (MVC). Computer image analysis (CIE94 color difference equation) was used to estimate the magnitude of the color difference between stained and unstained retina. Agents that had favourable staining characteristics underwent safety testing using a retinal pigment epithelium (ARPE19) and glial (MIO–M1) cell culture model . Cells were exposed to each agent and viability was assessed with a mitochondrial enzyme (MTT) assay, and fluorescent live–dead probe (ethidium homodimer–1 / calcein–AM). Techniques were tested on the following agents: alcian blue; diethyloxadicarbocyanine; evans blue; fast green; fluorescein; janus green; methylene blue; naphthol green; neutral red; procian (reactive) yellow; rose bengal; and trypan blue. Results: For most dyes the calculated color differences increased linearly with dye concentration, although some showed a more exponential relationship. Five agents showed favourable staining characteristics: evans blue, rose bengal, naphthol green, neutral red, and trypan blue (MVC 0.02, 0.01, 0.1, 0.002, 0.01% respectively). Safety testing of these five agents did not show toxicity, except in glial cells exposed to rose bengal. Relative to the negative control (saline), these showed a 48% reduction in viability using the MTT assay (p <0.001; t = 4.71; CI 30 to 75%), and qualitative damage on fluorescence microscopy. Conclusions: There are thousands of biological stains available and many of these may be more effective or safer than those currently used for retinal surgery. This study provides a means of screening potentially useful vital stains.

Keywords: retina • retinal glia • retinal pigment epithelium 
×
×

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.

×