Optimal preservation of the shark retina for ultrastructural analysis: an assessment of chemical, microwave, and high-pressure freezing fixation techniques

Microscopy Research and Technique
Blake K HarahushShaun P Collin

Abstract

Recent advances in microwave chemical fixation (MCF) and/or high pressure freezing (HPF) combined with transmission electron microscopy have resulted in superior ultrastructural detail in a variety of tissue types. To date, selachian tissue has been fixed and processed using only standard chemical fixation (CF) methods, and the resulting ultrastructure has been less than ideal. In this study, we compared the ultrastructure of the fragile retinal tissue from the brown banded bamboo shark, Chiloscyllium punctatum, obtained using CF, MCF, and HPF methods. For all fixation protocols, ultrastructural preservation was improved by keeping the tissue in oxygenated Ringer solution until the time of fixation. Both MCF and HPF produced superior retinal ultrastructure compared to conventional CF. Although HPF occasionally resulted in very high quality ultrastructure, microwave fixation was almost comparable, quicker and far more consistent.

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Citations

Apr 1, 2014·BMC Biology·Nagayasu NakanishiBernard M Degnan
Apr 22, 2015·Progress in Retinal and Eye Research·Tyler W Stradleigh, Andrew T Ishida

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