What are the Challenges in Single Molecule Fluorescence?
Despite its advantages, single molecule fluorescence also presents several challenges:
Photobleaching: Fluorescent molecules can lose their ability to emit light upon prolonged exposure to excitation light, limiting the duration of observations. Background Noise: Achieving a high signal-to-noise ratio is critical, as background fluorescence and detector noise can obscure single molecule signals. Complex Data Analysis: The rich and often noisy data obtained from SMF experiments require sophisticated analysis techniques and computational tools. Sample Preparation: Preparing samples for single molecule studies can be challenging, requiring precise control over concentration, purity, and immobilization of molecules.
