Single-cell electroporation for in vivo imaging of neuronal morphology and growth dynamics

Abstract

Single-cell electroporation (SCE) is a technique for acutely transfecting or dye-labeling individual neurons within intact living tissues. In addition to fl uorescently labeling neurons, SCE can be used to conduct cellautonomous studies of protein function by co-delivering fl uorophores with DNA, RNA, antisense constructs, peptides, proteins, or drugs. SCE involves inserting a thin glass pipette into neural tissue to restrict an electric fi eld and exposure to a solution of delivery compounds to an individual neuron at the pipette tip. Application of a brief electric pulse induces transient pores in the target cell and iontophoretic transfer of delivery compounds only to that cell. SCE is not limited to specifi c cell types and leaves no residual delivery agents. SCE has proven to be useful for in vivo fl uorescent imaging of neuronal morphology and connectivity and for conducting time-lapse imaging of structural changes due to growth and plasticity. Furthermore, "targeted SCE" allows selecting neurons based on connectivity, protein expression, activity patterns, or receptive fi eld properties. Overall, SCE offers a relatively simple and highly versatile alternative to transgenic approaches for acutely labeling or transfecting post-differentiated neurons.