gms | German Medical Science

22nd Annual Meeting of the German Retina Society

German Retina Society

26.06. - 27.06.2009, Berlin

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Flat mounted internal limiting membrane preparation

Meeting Abstract

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  • Ricarda G. Schumann - University Eye Clinic of Munich
  • R. Scheler - University Eye Clinic of Munich
  • M. M. Schaumberger - University Eye Clinic of Munich
  • C. Haritoglou - University Eye Clinic of Munich
  • A. Kampik - University Eye Clinic of Munich
  • A. Gandorfer - University Eye Clinic of Munich

German Retina Society. 22nd Annual Meeting of the German Retina Society. Berlin, 26.-27.06.2009. Düsseldorf: German Medical Science GMS Publishing House; 2009. DocRG2009-26

doi: 10.3205/09rg27, urn:nbn:de:0183-09rg274

This is the English version of the article.
The German version can be found at: http://www.egms.de/de/meetings/rg2009/09rg27.shtml

Published: June 29, 2009

© 2009 Schumann et al.
This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by-nc-nd/3.0/deed.en). You are free: to Share – to copy, distribute and transmit the work, provided the original author and source are credited.

Outline

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Purpose: In conventional microscopy, internal limiting membrane (ILM) specimens are usually prepared by series cross sectioning. Thus, analyses of cell density and cell distribution are limited. We used a new preparation technique with flat mounted ILM specimens of idiopathic macular holes (IMH) that allowed an evaluation of the whole membrane.

Methods: Fourty surgical specimens of the ILM were obtained from 40 eyes during pars plana vitrectomy for IMH. Specimens were placed into 2% paraformaldehyde and 0.1% glutaraldehyde for fixation and were prepared as wholemounts on glass slides for interference microscopy. The mounting medium 4’,6-diamidino-2-phenylindole (DAPI) was used to stain the cell nuclei.

Results: By flat mounted membrane specimens the total area of removed ILM was shown ranging from 3.2 mm2 to 22.6 mm2 with a mean of 12.0 mm2. Cell proliferation was found in all ILM specimens. The total number of cells ranged from 18 to 6914 with a mean of 1663. Cell distribution patterns varied between homogenous and inhomogenous with cell clusters. Cell density in specimens with homogenous cell distribution was 95 cells/mm2 compared to 365 cells/mm2 in cell clusters.

Conclusion: Preparation of flat mounted ILM specimens shows the whole membrane en face. The new preparation technique is suited for analyses of cell quantification and distribution. We first report on cell proliferation in 100% of all evaluated eyes with IMH. Cell differentiation and definite allocation of cell proliferation to the vitreal side or the retinal side of the ILM is not possible without further investigation.