Versatile Scientific Single-Molecule Fluorescence Microscopy Experiments for Advanced Chemistry Students

High Resolution Image Download MS PowerPoint Slide Although chemistry conceptionally is a molecular science, experiments are rarely performed on individual molecules but rather are typically carried out with an ensemble. Advanced chemistry students often already have considerable expertise in bulk spectroscopic methods. In order to introduce them to methods of single-molecule detection and chemistry, here we present a versatile, potentially low-cost, and easy-to-set-up course in single-molecule fluorescence microscopy. Using total internal reflection fluorescence microscopy (TIRFM) to image the fluorescence of single terrylene molecules in a solid p -terphenyl matrix, students learn not only to visualize single emitters but also to extract important quantitative photophysical data such as the emitted count rate, the photobleaching quantum yield, and the optical resolution or even to analyze chemical kinetics on the single-molecule level. Besides acquiring advanced experimental skills such as working with lasers and handling optical setups, the primary focus of this experiment lies on the evaluation and interpretation of the data collected, which includes the assessment of the setup and the critical interpretation of the results with reference to existing scientific models.