The field of Oncology is largely concerned with elucidating how cancers develop and how best to develop therapies to treat them. Oncologists commonly employ microscopy not only to identify and classify cancer cells, but also to identify the fragmented nuclei of apoptotic cells treated with a chemotherapeutic drug, or to visualize the binding and subsequent internalization and trafficking of therapeutic monoclonal antibodies. Amnis imaging flow cytometry is ideally suited to these types of studies, and gives the added benefit of combining immunophenotyping with quantitative morphometric measurements that can be applied to identifying rare sub-populations of potentially metastatic cells, including circulating tumor cells (CTC).
The FlowSight imaging flow cytometer offers population resolution using gross fluorescence measurements comparable to that of a PMT-based BD LSRFortessa CFC, in that cell division can be tracked by CellTrace Violet dilution and then subdivided into constitutive cell cycle phase using the PI/pH3 signals (figure 1 and figure 2). However, the imaging capabilities of the FlowSight allow the further subdivision of the mitotic phase into the three major stages using simple nuclear channel mask adaptations (figure 2) combined with just two morphometric measurements (figure 3). It is now possible to specifically look at compounds that affect the cell cycle down to the stages of mitosis. The imaging capability of the FlowSight, in combination with its speed, were essential to this study.