Nuclease‐Resistant L‐DNA Tension Probes Enable Long‐Term Force Mapping of Single Cells and Cell Consortia

Abstract DNA‐based tension probes with precisely programmable force responses provide important insights into cellular mechanosensing. However, their degradability in cell culture limits their use for long‐term imaging, for instance, when cells migrate, divide, and differentiate. This is a critical limitation for providing insights into mechanobiology for these longer‐term processes. Here, we present DNA‐based tension probes that are entirely designed based on the stereoisomer of biological D‐DNA, i.e., L‐DNA. We demonstrate that L‐DNA tension probes are essentially indestructible by nucleases and provide days‐long imaging without significant loss in image quality. We also show their superiority already for short imaging times commonly used for classical D‐DNA tension probes. We showcase the potential of these resilient probes to image minute movements, and for generating long term force maps of single cells and of collectively migrating cell populations.