Enables quantitation of neurite-specific metrics, such as neurite length, branch points and cell body clusters in co-cultures of neurons or neuronal cell lines with astrocytes
- Efficiently infects multiple neuronal cell types, including cell lines, primary and stem cell-derived cultures
- Synapsin-promoter driven expression of mKate2, a red fluorescent protein, ensures minimal expression in non-neuronal cell types
- Reagent is non-perturbing to neuronal biology
- Each vial contains 0.45mL of reagent, sufficient for at least 1 x 96-well plate
Schematic of Labeling Protocol
Example Images of NeuroLight Red Expression
UL and LL: Primary rat hippocampal and cortical neurons (GlobalStem) in co-culture with rat astrocytes (IncuCyte® NeuroPrime rAstrocytes, Essen Cat No 4586) at day 5 and 7 in vitro, respectively. UR: iPSC neurons (iCells, Cellular Dynamics International) were grown with iCell Astrocytes (Cellular Dynamics International). LR: Rat dorsal root ganglion neurons (DRG, Lonza) at day 7 in vitro.
- 3rd generation HIV-based, VSV-G pseudotyped lentiviral particles encoding a red fluorescent protein (mKate2).
- Promoter: Synapsin
- Spectral Properties: Ex (max): 588 nm; Em (max): 633 nm
Third generation lentiviral-based vectors are commonly used to transfer genetic information to cells for gene therapy and/or research purposes. The IncuCyte® NeuroLight lentiviral-based reagent has been specially designed to efficiently transduce multiple neuronal cell types with low toxicity. The NeuroLight Red lentivirus encodes a red fluorescent protein (mKate2) driven off a synapsin promoter to strengthen neuronal expression and minimize non-neuronal crossover. 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 reagent provide an integrated solution for kinetically measuring neurite dynamics in vitro.