Dyes providing increased sensitivity in flow-cytometric dye-efflux assays for multidrug resistance

In an effort to improve detection of P-glycoprotein-mediated multidrug resistance (mdr), several dyes were compared to rhodamine 123 (R123) in efflux assays. Two dyes (SY-38 and SY-3150) were found that provided better sensitivity. These dyes were effluxed by a cell line known to be mdr-positive (P388/R84) but not by an mdr-negative cell line (P388). Efflux was blocked by both verapamil and cyclosporine A. Efflux from KG1a cells was less than from P388/R84, just as has been seen with R123 and daunomycin. In further experiments, a model system was used to demonstrate two-color immunofluorescence plus efflux measurements. This was done using a sequential staining method. A procedure was devised that, at least for this model system, allowed single-step staining with both dye and antibody. The sensitivity of the efflux measurement was slightly compromised by using this one-step staining method.