Wide field time-domain FLIM

Wide field time-domain FLIM implementations... 

To use this type of microscope for FLIM, the camera has to be able to measure time resolved. Typically, a gain-modulated image intensifier is used for this, although recently the use of a modulated CCD camera for this has been reported [22,23]. The light source has to be intensity-modulated. This can be pulsed, for time-domain FLIM, or continuously, for frequency-domain FLIM. If a laser is used for excitation, the beam will have to be expanded to illuminate not a single point in the specimen but the entire field of view in order to obtain wide-field images. For references to wide-field FLIM instrumentation see Table 1, A1 and Bl. 

Lifetime imaging can be implemented both in wide field and in scanning microscopes such as confocal microscopes and two-photon excitation microscopes. The most common implementations in time-domain fluorescence lifetime imaging microscopy (FLIM) are based on TCSPC [8, 9] and time-gating (TG) [2, 10],... 

A more common approach to time domain wide field FLIM is based on a time-gated image intensifier MCP in combination with a CCD camera (see Fig. 3.8). After every excitation pulse the gated... 

---