The IncuCyte® NeuroLight Red Reagent is a third generation lentiviral-based vector encoded with a red fluorescent protein (mKate2) driven off of a synapsin promoter designed to efficiently transduce multiple neuronal cell types. Our extensive validation experiments have shown that expression of this red fluorescent protein does not negatively alter functional cell biology (e.g. morphology, neurite outgrowth, and neurite branching) of neurons in co-culture with astrocytes. In combination, the IncuCyte® Live-Cell Analysis System and NeuroLight Red Reagent provide an integrated solution for kinetically measuring neurite dynamics in vitro.