An imaging system includes a platform for placement of a sample or an animal to be imaged, and at least one excitation light source for irradiating the sample or animal to stimulate an emission at a plurality of different center wavelengths. An acousto-optic tunable filter (AOTF) is provided that includes a piezoelectric transducer crystal for emitting an acoustic wave having a ground electrode disposed on one side of the piezoelectric crystal. A patterned electrode layer is disposed on a side of the piezoelectric crystal opposite the ground electrode. The patterned electrode layer includes a continuous region proximate to its center and a discontinuous region, a pattern in the discontinuous region comprising a plurality of spaced apart features electrically connected to the continuous region, and an AO interaction crystal receiving the acoustic wave attached to the ground electrode or the patterned electrode layer.