Protocol for subcellular targeted microthermal protein damage in cells cultivated on plasmonic nanosilver-modified surfaces

Abstract Despite proteotoxic stress and heat shock are implicated in diverse pathologies, currently no methodology to inflict defined, subcellular thermal damage exists. Here, we present a protocol for such a single-cell method compatible with laser-scanning microscopes, adopting the plasmon resonance principle. The method is based on modified microscopic cell culture plates, pre-coated by a layer of anisotropic silver NPs allowing excitation through targeted irradiation by conventional lasers used in the laser scanning microscopes and allowing controllable heating. Dose-defined heat causes protein damage in subcellular compartments, rapid heat-shock chaperones recruitment and stress signalling, thereby allowing unprecedented spatiotemporal analysis of thermal damage with broad applicability in biomedicine.