Direct single-molecule detection and super-resolution imaging with a low-cost portable smartphone-based microscope

Detecting single molecules, which represents the ultimate level of sensitivity, is typically achieved with research-grade equipment. Here we present a low-cost, portable smartphone-based fluorescence microscope capable of detecting single-molecule fluorescence directly, i.e., without the need for any signal amplification. The setup leverages the image sensors and data handling capacity of mass-produced smartphones, it is adaptable to different smartphones and capable of detecting single molecules across the visible spectral range. We showcase this capability through single-molecule measurements on DNA origami models and super-resolution microscopy of cells by single-molecule localization microscopy. Last, we illustrate its potential as a point-of-care (POC) device by implementing a single-molecule bioassay for RNA detection. This development paves the way for innovative applications of massively distributed or personalized assays with single-molecule sensitivity in various fields such as digital bioassays, POC diagnostics, field expeditions, STEM outreach, and life science education.