Preparation of frozen sections for analysis.
The analysis of frozen tissue by antibodies can be accomplished by the quick freezing of a small tissue sample in liquid nitrogen. Super-cooled isopentane can also be used to further the preservation process. Freezing preserves the available proteins in a near-native state for their identification by antibodies raised against naturally folded proteins. The tissues are sectioned onto charged glass slides where they can be optimally fixed in weakly or non-denaturing solutions such as acetone or those that are alcohol-based. Following mild pretreatment steps to allow for antibody use with low background, (the endogenous peroxidase enzyme or oxidative compounds quenched in a hydrogen peroxide solution and available charged sites blocked by incubation in a normal serum solution) the sections are ready for antigen detection.
Bratthauer
Gary L
GL
Department of Gynecologic and Breast Pathology, Armed Forces Institute of Pathology, Washington, DC, USA.
D000818:Animals; D000941:Antigens / Q000032:analysis; D005629:Frozen Sections / Q000379:methods*; D006801:Humans; D009584:Nitrogen / Q000737:chemistry; D010420:Pentanes / Q000737:chemistry; D013997:Time Factors; D016707:Tissue Fixation / Q000379:methods
D000941:Antigens; D010420:Pentanes; D009584:Nitrogen; C067038:isopentane