Silanes to Silatranes: Robust Functionalization for Single‐Molecule Force Spectroscopy

Atomic force microscopy‐based single‐molecule force spectroscopy (SMFS) experiments require stable and reproducible functionalization of cantilevers and surfaces. Conventional silane‐based methods suffer from poor stability, drastically hindering their reproducibility and performance. Here, we present an ethanol‐based aminopropyl‐silatrane functionalization protocol that achieves superior performance in a similar process timeframe. In particular, combining this approach with bacterial adhesin coupling, we demonstrated a significant twofold improvement in the fraction of attempted SMFS experiments yielding usable single‐molecule data. The method can be generally applied to silicon‐nitride and glass cantilevers and surfaces, including mechanically modified cantilevers optimized for fast time response, low stiffness, and low drift. Furthermore, we demonstrate a simple and cost‐efficient method to clean these functionalized cantilevers using common solvents and a benchtop UV‐ozone cleaner, allowing for refunctionalization and prolonged cantilever lifespans. Together, this allows for a reliable, reusable, and versatile functionalization protocol for high‐quality SMFS experiments.