Detection of ultrasound (US)-modulated fluorescence in turbid media is a challenge because of the low level of fluorescent light and the weak modulation of incoherent light. A very limited number of theoretical and experimental investigations have been performed, and this is, to our knowledge, the first demonstration of pulsed US-modulated fluorescence tomography. Experimental results show that the detected signal depends on the acoustic frequency and the fluorescent target’s size along the ultrasonic propagation axis. The modulation depth of the detected signal is greatest when the length of the object along the acoustic axis is an odd number of half wavelengths and is weakest when the object is an integer multiple of an acoustic wavelength. Images of a fluorescent tube embedded within a 22- by 13- by 30 mm scattering gel phantom (, ) with 1-, 1.5-, and 2 MHz frequency US are presented. The modulation depth of the detected signal changes by a factor of 5 depending on the relative size of the object and the frequency. The approach is also verified by some simple experiments in a nonscattering gel and using a theoretical model.