Author/Editor		Srebotnik-Kirbiš, I; Strojan-Fležar, M; Us-Krašovec, M
Title		MIB-1 immunostaining on cytological samples: a protocol without antigen retrieval
Type		članek
Source		Cytopathology
Vol. and No.		Letnik 15, št. 3
Publication year		2004
Volume		str. 154-9
Language		eng
Abstract		The aim of the study was to evaluate the effect of fixation procedures on MIB-1 immunostaining on microwave treated Papanicolaou-stained slides and tn establish protocol for MIB-1 immunostaining on cytological samples without microwave pre-treatment. Cytospins for immunostaining and nuclear suspension for DNA measurement were prepared from human breast cancer cell line (MCF-7). Following fixation, the cytospins were either stained by Papanicolaou, stored in methanol or air-dried. Antigen retrieval by microwave was used betore MIB-1 immunostaining only for Papanicolaou-stained cytospins. Air-dried cytospins and cytospins stored in methanol were immunostained without pre-treatment. The percentage ot MIB-1 positive cells was compared with the S phase fraction of MCF-7 cells calculated from DNA histograms. Variations in fixation procedures used before Papanicolaou staining had no influence on the percentage of MIB-1 positive cells. The difference between the percentage of the MIB-1-positive cells on microwave-treated Papanicolaou-stained cytospins and on methanolfixed cytospins without microwave pre-treatment was not significant. There was a strong correlation between the percentage of the MIB-1-positive cells and S phase fraction. Monoclonal antibody MIB-1 recognized Ki-G7 antigen in Papanicolaou-stained cytospins treated by microwave as well as in cytospins that were fixed and stored in methanol without microwave pre-treatment.
Descriptors		TUMOR CELLS, CULTURED CYTOLOGICAL TECHNIQUES IMMUNOHISTOCHEMISTRY ALCOHOL, METHYL FIXATIVES