We present two complementary fluorescence measurement systems designed for quantitative analysis of molecular dynamics. A custom Fluorescence Correlation Spectroscopy (FCS) setup based on a 488 nm laser diode and time-correlated single-photon counting (TCSPC) provides single-molecule sensitivity for determining molecular concentration, diffusion, and microenvironmental interactions within a confocal detection volume. The system was validated using Rhodamine 110 in aqueous and sucrose solutions, yielding diffusion times and focal volume values consistent with commercial FCS instruments.
Also, we introduce a streak camera–based fluorescence lifetime measurement system supported by a comprehensive calibration protocol that corrects spectral, temporal, and intensity non-uniformities using a single reference image. Implemented on a Hamamatsu streak camera, the method was validated with Rhodamine 110 and further demonstrated on BodiPy-PM, resolving multi-component decay dynamics through deconvolution with the instrument response function.