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Effect of microwave irradiation on antibody labeling efficiency when applied to ultrathin cryosections through fixed biological material
Author(s) -
Chicoine Linda,
Webster Paul
Publication year - 1998
Publication title -
microscopy research and technique
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.536
H-Index - 118
eISSN - 1097-0029
pISSN - 1059-910X
DOI - 10.1002/(sici)1097-0029(19980701)42:1<24::aid-jemt4>3.0.co;2-r
Subject(s) - immunolabeling , polyclonal antibodies , antigen retrieval , antibody , antigen , chemistry , irradiation , biophysics , primary and secondary antibodies , microwave , microbiology and biotechnology , immunohistochemistry , biology , immunology , physics , quantum mechanics , nuclear physics
To study the effect of microwaves on immunolabeling, ultrathin cryosections or diluted antibodies were irradiated prior to antibody application, and gold labeling was quantified. In addition, affinity purified, polyclonal antibodies and protein A‐gold were applied to ultrathin cryosections of aldehyde‐fixed material in the presence and absence of microwaves. Amylase, a soluble protein secreted by pancreatic acinar cells, MHC class II, an integral membrane protein, and 3‐(2,4‐dinitroanilino)‐3‐amino‐N‐methyldipropylamine (DAMP), an exogenously added antigen, were localized with monospecific antibodies. Each was chosen for their contrasting subcellular characteristics. Results demonstrated that for some antigens, antibody labeling efficiency was quantitatively improved by microwave irradiation of sections prior to antibody application. Irradiation of diluted antibodies prior to their application also resulted in improved labeling. In contrast, the results obtained using rapid immunolabeling protocols in the presence of microwaves resulted in labeling levels similar to those obtained in the absence of microwaves. We conclude that microwave irradiation can improve the labeling efficiency of some antibodies. However, improvements in labeling density are dependent on the antigen under study and on variable irradiation times, unique to each antibody. This suggests that the routine use of microwaves to reduce incubation times may not be a viable alternative to bench protocols. Microsc. Res. Tech. 42:24–32, 1998. © 1998 Wiley‐Liss, Inc.