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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.
Fluorescence lifetime overlay of dyes Hoechst 33342 and Rhodamin 123


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

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