Comparative analysis of two immunohistochemical methods for antigen retrieval in the optical lobe of the honeybee Apis mellifera: Myosin-v assay

• Main Authors: LUCIANA KAREN CALÁBRIA, RENATA ROLAND TEIXEIRA, SYBELLI MAGDA COELHO GONÇALVES, ANDREIA BARCELOS PASSOS LIMA, ANA ALICE DINIZ DOS SANTOS, ANTÔNIO ROBERTO MARTINS, FOUED SALMEN ESPINDOLA
• Format: Article
• Language: English
• Published: BMC 2010-01-01
• Series: Biological Research
• Subjects: antigen retrieval; microwave oven; steam cooker
• Online Access: http://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0716-97602010000100002

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.