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An antibody panel for immunohistochemical analysis of the retina in Davidson's-fixed, paraffin-embedded eyes of rats.
- Source :
-
Experimental and toxicologic pathology : official journal of the Gesellschaft fur Toxikologische Pathologie [Exp Toxicol Pathol] 2009 Mar; Vol. 61 (2), pp. 91-100. Date of Electronic Publication: 2008 Aug 20. - Publication Year :
- 2009
-
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.
- Subjects :
- Animals
Apoptosis
Biomarkers analysis
Caspase 3 immunology
Cell Proliferation
Eye chemistry
Eye immunology
Glial Fibrillary Acidic Protein immunology
Glutamate-Ammonia Ligase immunology
Histones immunology
Male
Rats
Rats, Long-Evans
Rhodopsin immunology
Sensitivity and Specificity
Staining and Labeling
Synaptophysin immunology
Antibodies, Monoclonal immunology
Eye pathology
Immunohistochemistry methods
Paraffin Embedding
Subjects
Details
- Language :
- English
- ISSN :
- 1618-1433
- Volume :
- 61
- Issue :
- 2
- Database :
- MEDLINE
- Journal :
- Experimental and toxicologic pathology : official journal of the Gesellschaft fur Toxikologische Pathologie
- Publication Type :
- Academic Journal
- Accession number :
- 18718747
- Full Text :
- https://doi.org/10.1016/j.etp.2008.06.005