Starting from a discussion of the peculiarities of high-resolution low-level optical signal recording, this article describes the recording process of classic TCSPC and its extension, multidimensional TCSPC. It shows why TCSPC reaches a time resolution, sensitivity, and photon efficiency beyond the reach of any other optical signal recording technique. The article then passes to the general features of the bh TCSPC technique: Outstanding time resolution, outstanding timing stability, and a unsurpassed variety of multi-dimensional recording principles. These features are demonstrated on examples of high-resolution fluorescence decay recording, multi-detector operation and laser multiplexing, simultaneous fluorescence and phosphorescence recording, parameter-tag recording, FLIM, multi-wavelength FLIM, spatial and temporal mosaic FLIM, and simultaneous FLIM/PLIM.
28 pages, 36 figures, 50 references
Keywords: TCSPC, FLIM, PLIM, FRET, FLIO, FCS, Metabolic Imaging, Molecular Imaging