The percentage of apoptotic cells in a sample can be accurately and objectively determined using OTMIAS® and the O-STANDARDS™ specific for the apoptosis assay after staining the sections for Cleaved Caspase-3.
In the top figure on the left, cultures of cell lines were treated with an apoptosis-inducing drug at different concentrations and for different time periods. Cells were trypsinized and suspended in culture media, then a sample of each was taken for apoptosis determination by Flow Cytometry after annexin-V-FLUOS and propedium iodide double staining . The remaining cell suspension from each culture was fixed in 10% buffered formalin and used to prepare cytologic preparations (CytoPreps) on glass slides (whole cells). These CytoPrep and O-STANDARD slides were stained for Cleaved Caspase-3 utilizing the immunoperoxidase method, then the percentage of apoptotic cells was determined utilizing OTMIAS® and standard slides. The graph shows the correlation between the percentage of apoptotic cells for the various cell cultures determined utilizing OTMIAS® and Flow Cytometry. Similar results can be obtained using sections of formalin fixed and paraffin embedded tissues or cells.
OTMIAS® gives the results of the apoptosis assay as a bar graph with the percentage of apoptotic cells (bottom figure on the right).
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