Simultaneous Topography, Stiffness, and Surface Potential Mapping of Individual Cells Using Scanning Ion Conductance Microscopy

Developing multifunctional imaging techniques to acquire multiple parameters simultaneously is critical for advancing single-cell analysis. In this study, we present a novel approach using Scanning Ion Conductance Microscopy (SICM) to simultaneously map the topography, stiffness, and surface potential of live and fixed cells. Using a double-barrel nanopipette, we can measure both ionic current and surface potential signals near the location of the nanopipette tip. By optimizing imaging conditions and data processing methods, we successfully acquired these three types of images concurrently We validated the technique on HEK and CAL27 cells, providing high-resolution images that capture cell membranes' mechanical and electrostatic properties. This approach offers a comprehensive tool for understanding the interplay between cellular morphology, mechanics, and electrostatics, facilitating more detailed insights into cellular behavior and response to stimuli.