Seminarium Optyczne

Single-molecule microscopy and spectroscopy are advanced research techniques that enable the real-time tracking and analysis of individual molecules. These methods provide detailed insights into molecular dynamics, interactions, and structures, which cannot be achieved by studying phenomena at the ensemble level. Two techniques in this field, SOFI (Stochastic Optical Fluctuation Imaging) and defocused wide-field fluorescence microscopy, offer unique approaches for high-resolution molecular/nanoparticle analysis. SOFI is based on the statistical analysis of fluorescence signal fluctuations generated by single emitters. This allows for image reconstruction with a resolution beyond the diffraction limit of light, particularly for densely labeled samples. Defocused wide-field fluorescence microscopy involves imaging under deliberately defocused conditions. The emerging single-molecule patterns provide information about the orientation and structure of individual molecules. This technique is particularly suitable for studying rotational dynamics (e.g., molecular motors), allowing for the direct observation of even tens of individual molecules. Both methods are powerful tools in modern molecular research, providing extensive possibilities for analyzing structures and processes at the nanoscale level.