Magnetic Nano-Actuation Platform for Remote and Controllable Mechanostimulation of Schwann Cells with Real-Time Confocal Imaging.

Mechanical forces critically regulate cellular behavior, yet many existing methods of mechanical stimulation rely on direct physical contact or artificially engineered extracellular environments, thereby limiting their flexibility in dynamic live-cell studies. Here, we present a magnetic nano-actuation platform for remote, non-contact, and controllable mechanostimulation of Schwann cells in vitro. This integrated setup enables remote magnetic stimulation and synchronized live-cell imaging in the same experimental session. This manuscript describes the preparation of fluorescent superparamagnetic nanoparticles (SPIONs), optionally actin-targeting functionalized SPIONs (f-SPIONs), the construction and calibration of a microscope-compatible electromagnetic stimulation device, the estimation of magnetic forces at the single-particle and single-cell levels, and the integration of magnetic actuation with real-time confocal imaging. Schwann cells are highly mechanosensitive glial cells that play essential roles in the development, maintenance, and repair of peripheral nerves. Real-time monitoring of their responses to mechanical stimulation is crucial for understanding how mechanical forces influence cytoskeletal organization and cellular behavior. This platform provides a reproducible workflow for studying Schwann cell mechanobiology and may be adapted to other mechanically responsive cell types and multicellular systems.