Septal penetration of high energy photons may degrade 123I images obtained with a low energy collimator. We evaluated the physical characteristics of a low energy, high resolution (LEHR) collimator, special LEHR (SLEHR) collimator, and medium energy collimator for 123I studies. The cross-talk of 99mTc and 123I into the 201Tl window was also examined. Sensitivity and spatial resolution were measured with each collimator. Point sources of 99mTc and 123I were imaged at various source-collimator distances using multiple energy windows, and the effects of collimator choice on energy spectrum and spatial distribution of photopeak counts were assessed. For 99mTc, both sensitivity and resolution were similar with the low energy collimators, and higher sensitivity and lower resolution were observed with the medium energy collimator. For 123I, the full width at tenth maximum was larger for 123I than for 99mTc when using the LEHR collimator. Acquisitions with multiple energy windows revealed severe degradation due to septal penetration in imaging 123I with the LEHR collimator, especially at short distances. The degradation was reduced with the SLEHR collimator and further with the medium energy collimator. In both 99mTc and 123I imaging, cross-talk into the 201Tl window was larger at shorter distances and the largest with the LEHR collimator. In conclusion, variation in collimator geometry causes differences in the effect of septal penetration on 123I images and in cross-talk into the 201Tl window. The SLEHR collimator may be suitable for use in high resolution 123I imaging and simultaneous 99mTc/201Tl imaging. Use of the medium energy collimator appears to be preferable in quantitative 123I studies.