Live-cell SICM imaging: An Introductory Guide for New Users

High-resolution, minimally invasive imaging of live cells is essential for investigating cellular morphology and its dynamic changes. Among available approaches, Scanning Ion Conductance Microscopy (SICM) offers a unique combination of precise topographic imaging down to nanoscale with experimental conditions that preserve true livecell behavior. Here, we present a practical guide based on our own experience and experiments, intended to be a resource to help first-time SICM users, covering critical aspects from instrumentation, probe characterization, to cell preparation and morphometric data extraction and presentation, to accelerate their implementation, learning and to gain confidence in exploring live-cell structure and dynamics.