title: Comparative analysis of two immunohistochemical methods for antigen retrieval in the optical lobe of the honeybee Apis mellifera: myosin-V assay.
abstract: The present study compared two heating methods currently used for antigen retrieval (AR) immunostaining: the microwave oven and the steam cooker. Myosin-V, a molecular motor involved in vesicle transport, was used as a neuronal marker in honeybee Apis mellifera brains fixed in formalin. Overall, the steam cooker showed the most satisfactory AR results. At 100 ºC, tissue morphology was maintained and revealed epitope recovery, while evaporation of the AR solution was markedly reduced; this is important for stabilizing the sodium citrate molarity of the AR buffer and reducing background effects. Standardization of heat-mediated AR of formalin-fixed and paraffin-embedded tissue sections results in more reliable immunostaining of the honeybee brain.
authors.affiliation: Instituto de Genética e Bioquímica, Universidade Federal de Uberlândia, Uberlândia, MG 38400-902, Brazil.
authors.forename: Luciana Karen
authors.initials: LK
authors.lastname: Calábria
authors.forename: Renata Roland
authors.initials: RR
authors.lastname: Teixeira
authors.forename: Sybelli Magda
authors.initials: SM
authors.lastname: Coelho Gonçalves
authors.forename: Andreia Barcelos
authors.initials: AB
authors.lastname: Passos Lima
authors.forename: Ana Alice Diniz Dos
authors.initials: AA
authors.lastname: Santos
authors.forename: Antônio Roberto
authors.initials: AR
authors.lastname: Martins
authors.forename: Foued Salmen
authors.initials: FS
authors.lastname: Espindola
mesh_terms: D000818:Animals; D000941:Antigens / Q000032:analysis* / Q000276:immunology; D001516:Bees / Q000276:immunology*; D006361:Heating; D007150:Immunohistochemistry / Q000379:methods*; D008872:Microwaves; D024701:Myosin Type V / Q000032:analysis*; D009899:Optic Lobe, Nonmammalian / Q000737:chemistry*; D016612:Paraffin Embedding; D013194:Staining and Labeling
chemical_list: D000941:Antigens; D024701:Myosin Type V