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MIB-1 immunostaining on cytological samples: a protocol without antigen retrieval.
- Source :
-
Cytopathology : official journal of the British Society for Clinical Cytology [Cytopathology] 2004 Jun; Vol. 15 (3), pp. 154-9. - Publication Year :
- 2004
-
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 to 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 before MIB-1 immunostaining only for Papanicolaou-stained cytospins. Air-dried cytospins and cytospins stored in methanol were immunostained without pre-treatment. The percentage of 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 methanol-fixed 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-67 antigen in Papanicolaou-stained cytospins treated by microwave as well as in cytospins that were fixed and stored in methanol without microwave pre-treatment.
Details
- Language :
- English
- ISSN :
- 0956-5507
- Volume :
- 15
- Issue :
- 3
- Database :
- MEDLINE
- Journal :
- Cytopathology : official journal of the British Society for Clinical Cytology
- Publication Type :
- Academic Journal
- Accession number :
- 15165272
- Full Text :
- https://doi.org/10.1111/j.1365-2303.2004.00146.x