Shrinkage in preparatory steps for SEM. A study on rabbit corneal endothelium

Albrecht Von Graefes Archiv Für Klinische Und Experimentelle Ophthalmologie. Albrecht Von Graefe's Archive for Clinical and Experimental Ophthalmology
O A JensenH Laursen

Abstract

Since specular microscopy of the cornea offers the opportunity to observe and measure cells in vivo without any outside interference this method forms an unrivalled basis for estimation of tissue shrinkage during various preparatory methods. Therefore a study was performed with the purpose of evaluating the degree of artifacts in each preparatory step from the living tissue "in vivo" to the final SEM specimen. The study was performed on rabbit corneas, the endothelium serving as measuring target. The in vivo state was recorded by specular microscopy. Unfixed corneas were studied by light microscopy unstained and stained by alizarin red S or silver nitrate. Fixation was performed intracamerally with 1.5% glutaraldehyde (Gla) by a pH, osmolarity, viscosity and intraocular pressure identical with the physiological values of rabbit eyes. Fixation was completed by immersion in 2.5% Gla for 1/2 h. Gla-fixed corneas were evaluated as above before osmification. Dehydration was performed either by graded acetone, by acetone in a gradient-free system, both followed by critical point drying (CPD). At all steps cells were counted using the same reference frame. The number of cells/mm2 was estimated and statistical analysis showed a shrinka...Continue Reading

References

Jun 1, 1978·Acta Ophthalmologica·S Sperling
Jul 1, 1978·The American Journal of Anatomy·S Billings-GagliardiG B Schneider
Nov 1, 1978·Archives of Ophthalmology·A LauleC Hanna
Dec 1, 1978·The British Journal of Ophthalmology·G D SturrockN S Rice
Jun 1, 1976·Experimental Eye Research·R A LaingH M Leibowitz
Oct 1, 1977·American Journal of Ophthalmology·W L BlackwellH E Kaufman

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Citations

Jan 1, 1986·Graefe's Archive for Clinical and Experimental Ophthalmology = Albrecht Von Graefes Archiv Für Klinische Und Experimentelle Ophthalmologie·S Sperling
Jan 1, 1994·Progress in Histochemistry and Cytochemistry·S R Gordon
May 24, 2011·Nanomedicine : Nanotechnology, Biology, and Medicine·Rebecca A DragovicIan L Sargent
Aug 26, 2014·Journal of Extracellular Vesicles·Maria T AatonenPia R-M Siljander
Jan 19, 2018·Scientific Reports·Lígia Nunes de Morais RibeiroEneida de Paula
Aug 1, 1984·Die Naturwissenschaften·G Ruhenstroth-BauerS Vogl

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