OTMIAS® enables investigators to obtain accurate and objective S-Phase Fraction (SPF) values in sections of formalin fixed and paraffin embedded tissues or cells after staining for Cyclin A using the immunoperoxidase method (IHC).
In the figure on the right, cultures of various cell lines were trypsinized, suspended in culture media, then a sample of each was taken for SPF determination by Flow Cytometry (the gold standard). The remaining cell suspension from each cell culture was fixed in 10% buffered formalin and used to prepare cytologic preparations (CytoPrep) on glass slides (whole cells), or to prepare paraffin blocks. CytoPrep slides and 4- micron sections of the paraffin blocks were stained for Cyclin A utilizing IHC, then the SPF was determined utilizing OTMIAS® with the appropriate O-STANDARDS™ as controls for calibration. The graph shows the correlation between SPF for the various cell lines determined by OTMIAS® compared to Flow Cytometry.
OTMIAS® gives the results of the proliferative assay as a bar graph with the percentage of cells in the s-phase of the cell cycle (bottom figure on the left).
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