Protocol for optogenetic stimulation of cells under physical confinement.
Abstract:
Optogenetic approaches enable spatiotemporal control of signaling proteins, yet their integration with microfluidic assays to study confined cell migration remains challenging. Herein, we present a protocol for the optogenetic activation of PI3K/Akt signaling in confined cells. We detail procedures for applying stimulation to induce localized Akt activation at the cell's leading edge. This protocol enables real-time manipulation of subcellular signaling dynamics during confined migration. For additional information on the use of this protocol, please refer to Lee et al.1.