Rapid multiple immunocytochemical staining method using microwave irradiation for intraoperative cytology.
OBJECTIVE
To develop a more rapid and accurate staining procedure for intraoperative cytology through an assessment of a rapid multiple immunocytochemical staining method using microwave irradiation.


METHODS
A preliminary test of a triple immunostaining method using microwave irradiation was performed to determine optimal incubating conditions for primary antibodies against MOC-31, BerEP4 and carcinoembryonic antigen. The test samples were adenocarcinoma cells obtained by washings from 10 resected colorectal cancer specimens. Adenocarcinoma cells in peritoneal washings diagnosed by cytologic examination from 8 colorectal cancer patients were retrospectively examined using the previously determined optimal incubating conditions.


RESULTS
High rates of positive cells (83-87%) with intense immunoreactions were obtained using a rapid process with primary antibody concentrations that were 2 times higher than those used for standard incubation at room temperature (82-86%). The staining under these conditions was completed within 19 minutes. Adenocarcinoma cells in the peritoneal washings were also intensely stained under these conditions.


CONCLUSIONS
Using our microwave method, the processing time was dramatically shortened, and intense and sensitive immunoreactions were obtained. The present method is useful for the rapid and accurate diagnosis of intraoperative cytology.
Division of Biomedical Imaging Research, Graduate School of Medicine, Juntendo University School of Medicine, Tokyo, Japan. fullhat@juntendo.ac.jp
Atsushi
A
Furuhata
Noriyoshi
N
Sueyoshi
Hidetake
H
Kurihara
Kazuhiro
K
Sakamoto
Toshiki
T
Kamano
Okio
O
Hino
D000230:Adenocarcinoma / Q000378:metabolism / Q000473:pathology / Q000601:surgery; D014408:Biomarkers, Tumor / Q000378:metabolism; D015179:Colorectal Neoplasms / Q000378:metabolism / Q000473:pathology / Q000601:surgery; D003581:Cytodiagnosis / Q000379:methods*; D006801:Humans; D007150:Immunohistochemistry / Q000379:methods*; D007432:Intraoperative Period; D008872:Microwaves; D010533:Peritoneal Lavage; D012189:Retrospective Studies; D013194:Staining and Labeling / Q000379:methods
D014408:Biomarkers, Tumor