The mechanical failure of polymers remains challenging to understand and predict, as it often involves highly localised phenomena that cannot be probed with bulk characterisation techniques. Here, we present a generalisable protocol based on optical microscopy, tensile testing, and image processing that permits the spatially resolved interrogation of mechanical deformation at the molecular level around defects in mechanophore-containing polymers. The approach can be applied to a broad range of polymeric materials, mechanophores, and deformation scenarios.