2-GHz frequency-domain fluorometer

Abstract

We developed a frequency-domain fluorometer which operates from 4 to 2000 MHz. The modulated excitation is provided by the harmonic content of a laser pulse train (3.76 MHz, 5 ps) from a synchronously pumped and cavity dumped dye laser. The phase angle and modulation of the emission are measured with a microchannel-plate photomultiplier (PMT). Cross-correlation detection is performed outside the PMT. The high-frequency signals for cross correlation were obtained by multiplication of the output from a 500-MHz frequency synthesizer. The performance was verified in several ways, including measurement of known time delays and examination of standard fluorophores. The detector displayed no detectable color effect, with the 300–600-nm difference being less than 5 ps. The precision of the measurements is adequate to detect differences of 20 ps for decay times of 500 ps. A correlation time of 53 ps was found for indole in water at 20 °C. The shortest correlation time we measured was 15 ps for indole in methanol/water (75/25) at 40 °C. Also, the 2-GHz data reveal the time-dependent ((t)1/2) terms found in the presence of collisional quenching. The degree of random error is about 0.3° of phase and 0.005 in modulation throughout the frequency range.