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Biophysical Journal 20: 335-342 (1977)
© 1977 the Biophysical Society

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A phase-shift fluorometer using a laser and a transverse electrooptic modulator for subnanosecond lifetime measurements.

I Salmeen and L Rimai

ABSTRACT

We described a simple phase-shift fluorometer using continuous laser excitation. The laser enables the use of a transverse mode electrooptic modulator with a half-wave retardation voltage of about 200 V (in contrast to many kilovolts of longitudinal modulators) at frequencies up to 100 MHz. The modulated fluorescence signal is detected, after passing through a double monochromator, by a photomultiplier tube feeding a radio frequency (RF) tuned amplifier. THE RF phase is then determined by phase-sensitive detection using a double balanced mixer with the reference obtained from a PIN photodiode-turned amplifier combination which detects light split off from the main exciting beam. The laser and double monochromator allow the observation of modulated Raman solvent and Rayleigh scatterin, which are convenient for determining the zero reference phase.






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Copyright © 1977 by the Biophysical Society.