1. Labeling: Cells or tissues are first labeled with a fluorescent dye or protein to visualize the molecules of interest. 2. Photobleaching: A high-intensity laser beam is focused on a specific region, causing irreversible bleaching of the fluorescent molecules in that area. 3. Recovery Monitoring: Post-bleaching, the movement of unbleached fluorescent molecules into the bleached area is monitored over time using time-lapse fluorescence microscopy. 4. Data Analysis: The recovery curve is analyzed to determine the diffusion coefficient and other kinetic parameters.
