Protocol for measuring the Young’s modulus of organoids using atomic force microscopy

Atomic force microscopy (AFM) is extensively applied to measure cell and tissue mechanics but lacks a standardized organoid stiffness assessment. Here, we present a protocol for quantifying the Young's modulus of organoids via AFM, combining force-curve analysis with an optimized probe. We describe steps for preparing organoids, OCT embedding, slicing, AFM detection, and force-curve analysis. By mechanically addressing gaps in organoids, this protocol improves reproducibility and expands the capabilities of biomechanical research.