AbstractNanopatterning methods based on particle lithography offer generic capabilities for high-throughput fabrication with thin film materials, such as organothiol and organosilane self-assembled monolayers (SAMs), polymer films, biological samples, and nanoparticles. Combining scanning probe microscopy with sample preparation based on approaches with particle lithography produces robust test platforms for ultrasensitive surface measurements. For example, nanopatterns of octadecyltrichlorosilane (OTS) can be prepared on surfaces of Si(111) using designed protocols of particle lithography combined with steps of either vapor deposition, immersion, or contact printing. Changing the physical approaches for applying molecules to masked surfaces produces nanostructures with designed shapes and thickness. Billions of nanostructures can be prepared using strategies for particle lithography, requiring only basic steps of mixing, heating, centrifuging, and drying. Arrays of exquisitely small and regular nanopatterns can be prepared with few defects and high reproducibility. For nanopatterns prepared with SAMs, the endgroups can be designed to spatially define the interfacial selectivity for adsorption of proteins, nanoparticles, or electrolessly deposited metals. Particle lithography has become a mature technique, with broad applicability for thin film materials. Images and measurements acquired with scanning probe microscopy will be described for samples prepared using particle lithography-based approaches.