A fluorescent-based assay for live cell, spatially resolved assessment of vesicular monoamine transporter 2-mediated neurotransmitter transport

► We report a method for spatially resolved analysis of VMAT2 mediated uptake in live cells. ► Uptake of the fluorescent substrate was measured in HEK293 cell lines stable expressing DAT and mCherry-VMAT2. ► Uptake into the mCherry positive compartment is sensitive to tetrabenazine. ► VMAT2 activity can be quantified by analyzing confocal images. ► We have developed an assay amenable to high content imaging that is low cost, uses no animals, and is flexible compared to radioactive assays.