An antibody panel for immunohistochemical analysis of the retina in Davidson's-fixed, paraffin-embedded eyes of rats.
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.
McKay
Jennifer S
JS
Department of Pathology, Safety Assessment, AstraZeneca, Cheshire, UK. jenny.mckay@astrazeneca.com
Steele
Samantha J
SJ
Ahmed
Gulshanara
G
Johnson
Elizabeth
E
Ratcliffe
Kerry
K
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D017209:Apoptosis; D015415:Biomarkers / Q000032:analysis; D053148:Caspase 3 / Q000276:immunology; D049109:Cell Proliferation; D005123:Eye / Q000737:chemistry / Q000276:immunology / Q000473:pathology*; D005904:Glial Fibrillary Acidic Protein / Q000276:immunology; D005974:Glutamate-Ammonia Ligase / Q000276:immunology; D006657:Histones / Q000276:immunology; D007150:Immunohistochemistry / Q000379:methods*; D008297:Male; D016612:Paraffin Embedding; D051381:Rats; D020318:Rats, Long-Evans; D012243:Rhodopsin / Q000276:immunology; D012680:Sensitivity and Specificity; D013194:Staining and Labeling; D016708:Synaptophysin / Q000276:immunology
D000911:Antibodies, Monoclonal; D015415:Biomarkers; D005904:Glial Fibrillary Acidic Protein; D006657:Histones; D016708:Synaptophysin; D012243:Rhodopsin; D053148:Caspase 3; D005974:Glutamate-Ammonia Ligase