Clinical Applications of Multispectral Imaging Flow Cytometry

The Image Stream is a flow cytometry-based image analysis platform that acquires up to 12 spatially correlated spectrally-separated images of cells in suspension at rates of up to 1000 cells/sec. By combining the high throughput and multiparameter capability of flow cytometry with the high image content information of microscopy it allows quantitative image analysis in immunophenotypically defined cell populations in statistically robust cell numbers. One area of its clinical application is in the study of cell signal transduction pathways for which the intracellular localization of signaling intermediaries correlate with activity. For example, activation of the nuclear factor-kappaB (NF kappa B) transcription factor complex is associated with the cytoplasmic to nuclear translocation of p65. To demonstrate this application, the nuclear translocation of p65 following receptor mediated and drug induced activation of NF kappa B was studied in human myeloid leukemia cells. TNF alpha -induced nuclear translocation of p65 was rapid and concentration-dependent, peaking at 30 min of exposure with maximum translocation achieved with concentrations above 5 ng/ml. Daunorubicin (DNR)-induced p65 translocation was concentration-dependent and correlated with DNR-induced apoptosis. The clinical context, the analysis approaches and results will be presented.