Abstract
Unlike most other tissues, the optimal fixative for preserving eye morphology is considered to be Davidson's fixative or modified Davidson's rather than formalin. However, the methodology for antibodies to be used in tissues fixed this way is not normally outlined in current antibody datasheets. Additionally, where eyes have been stored in Davidson's fixative, the efficacy of retrospective analysis of eye morphology by immunohistochemistry is largely unknown. The aim of this study was to compare a panel of six antibodies in both Davidson's-fixed and formalin-fixed pigmented and non-pigmented rat eyes, in order to provide optimal methods for future retinal immunohistochemical evaluation with image analysis. The antibodies evaluated were raised against rhodopsin, synaptophysin, glutamine synthetase, glial fibrillary acidic protein (GFAP), cleaved caspase-3 and phospho-histone H3 (PH3). Overall, the staining quality of these antibodies was found to be optimal in Davidson's compared to formalin-fixed tissues after a time period of up to 4 days in fixative. The methods outlined thus provide a platform for future detailed analysis of retinal pathology in Davidson's-fixed eyes.
References
Sep 1, 1992·Neurochemical Research·W T NortonC F Brosnan
Nov 1, 1987·The Journal of Histochemistry and Cytochemistry : Official Journal of the Histochemistry Society·D Hicks, C J Barnstable
Nov 1, 1984·The Journal of Comparative Neurology·R W Young
Jul 15, 1994·Brain Research. Developmental Brain Research·J P KapfhammerM E Schwab
Sep 1, 1995·Journal of Neuroscience Research·A Derouiche, T Rauen
Feb 13, 1999·Experimental Eye Research·N H ChongP J Luthert
Jun 23, 2001·Oncogene·F Hans, S Dimitrov
Jul 9, 2002·Eye·G P LewisS K Fisher
Oct 2, 2003·Brain Research. Developmental Brain Research·Rajesh K SharmaDianna A Johnson
Jan 6, 2006·Toxicologic Pathology·Tijana Mitić, Jennifer S McKay
Jan 13, 2006·The Journal of Histochemistry and Cytochemistry : Official Journal of the Histochemistry Society·Nobuo TeradaShinichi Ohno
Feb 7, 2006·Neuroscience·L P XueE-A Ling
Jul 28, 2007·Experimental Eye Research·Steven E WilsonFabricio W Medeiros
Citations
Aug 13, 2011·The Journal of Histochemistry and Cytochemistry : Official Journal of the Histochemistry Society·Glyn ChidlowRobert J Casson
Jul 14, 2010·Journal of Pharmacological and Toxicological Methods·William S RedfernJennifer S McKay
Jul 14, 2016·Current Eye Research·Ying Liu, Deepak P Edward
Dec 19, 2015·Toxicologic Pathology·Elke-Astrid AtzpodienPeter Maloca
Apr 25, 2018·Japanese Journal of Ophthalmology·Kazuhiro TokudaKoh-Hei Sonoda
Nov 10, 2017·Aging Cell·Yusuke IzutaKazuo Tsubota
Mar 13, 2019·Histochemistry and Cell Biology·Teresa MammoneJohn P M Wood
Oct 7, 2020·Veterinary Pathology·Vanessa VrolykMarie-Odile Benoit-Biancamano