Fluorescence lifetime overlay of dyes Hoechst 33342 and Rhodamin 123
| | Fluorescent proteins, molecular probes and various endogenous biomole-cules are excitable by ultrashort laser pulses. The fluorescence is character-ized by specific decay kinetics which can be used to distinguish between the fluorescent components and to obtain information on the microenvironment. TauMap® provides 3D fluorescence lifetime measurements (FLIM) for a spatial mapping of fluorescent molecules. The use of time correlated single photon counting (TCSPC) enables a temporal resolution of 50 ps / 250 ps with single photon sensitivity to detect even slight changes in fluorescence lifetime as well as second harmonic generation (SHG). The system is opti-mized for near infrared femtosecond laser pulses but can also be used in combination with UV / VIS ultrashort pulsed lasers.
| Fluorophore | Emission Wavelength
[nm] | Fluorescence Lifetime
[ns] | NAD(P)H | 450 - 470 | 0.3 (bound: 2) | Flavins | 530 | 5.2 (bound: <1) | Porphyrins | 580 – 720 | 10 - 15 | CFP | 475 / 500 | 1.3 / 3.8 | GFP (wild-type) | 510 | 2.7 / 3.1 | Collagen (SHG) | λ/2 (360-480) | 0 | |
