title: Antigen recovery and preservation using the microwave irradiation of biological samples for transmission electron microscopy analysis.
abstract: Most studies using microwave irradiation (MWI) for the preparation of tissue samples have reported an improvement in structural integrity. However, there have been few studies on the effect of microwave (MW) on antigen preservation during sample preparation prior to immunolocalization. This report documents our experience of specimen preparation using an automatic microwave apparatus to obtain antigen preservation and retrieval. We tested the effects of MW processing vs. conventional procedures on the morphology and antigenicity of two different tissues: the brain and mammary gland, whose chemical composition and anatomical organization are quite different. We chose to locate the transcription factor PPARβ/δ using immunocytochemistry on brain tissue sections from hamsters. Antigen retrieval protocols involving MWI were used to restore immunoreactivity. We also studied the efficiency of the ultrastructural immunolocalization of both PPARγ and caveolin-1 following MWI vs. conventional treatment, on mammary gland tissue from mice at 10 days of lactation. Our findings showed that the treatment of tissue samples with MWI, in the context of a process lasting just a few hours from fixation to immunolocalization, enabled similar, or even better, results than conventional protocols. The quantification of immunolabeling for cav-1 indicated an increase in density of up to three-fold in tissues processed in the microwave oven. Furthermore, MW treatment permitted the localization of PPARβ/δ in glutaraldehyde-fixed specimens, which was impossible in the absence of MWI. This study thus showed that techniques involving the use of microwaves could largely improve both ultrastructure and immunodetection.
authors: Institut National de la Recherche Agronomique (INRA), Unité de Recherche UR902 Nutrition et Régulation Lipidique des Fonctions Cérébrales, Jouy-en-Josas, France.
authors: Josiane
authors: J
authors: Aïoun
authors: Sophie
authors: S
authors: Chat
authors: Christian
authors: C
authors: Bordat
authors: Christine
authors: C
authors: Péchoux
mesh_terms: D000818:Animals; D000941:Antigens / Q000032:analysis*; D057205:Automation, Laboratory / Q000379:methods; D001921:Brain / Q000648:ultrastructure; D001923:Brain Chemistry; D051242:Caveolin 1 / Q000032:analysis; D006224:Cricetinae; D008321:Mammary Glands, Animal / Q000737:chemistry / Q000648:ultrastructure; D051379:Mice; D046529:Microscopy, Electron, Transmission / Q000379:methods*; D008872:Microwaves; D047494:PPAR delta / Q000032:analysis; D047495:PPAR gamma / Q000032:analysis; D048068:PPAR-beta / Q000032:analysis; D011309:Preservation, Biological / Q000379:methods*; D013048:Specimen Handling / Q000379:methods*
chemical_list: D000941:Antigens; D051242:Caveolin 1; D047494:PPAR delta; D047495:PPAR gamma; D048068:PPAR-beta
keywords: Antigen preservation; Antigen retrieval; Brain; Mammary gland; Microwave; Ultrastructural immunocytochemistry