Purpose: We compared the reproducibility of $^{201}Tl;and;^{99m}Tc$-sestamibi (MIBI) gated SPECT measurement of myocardial function using the Germano algorithm Materials and Methods: Gated SPECT acquisition was repeated in the same position in 30 patients who received $^{201}Tl$ and in 26 who received $^{99m}Tc$-MIBI. The quantification of end-diastolic volume (EDV), end-systolic volume (ESV), and ejection fraction (EF) on $^{201}Tl;and;^{99m}Tc$-MIBI gated SPECT was processed independently using Cedars quantitative gated SPECT software. The reproducibility of the assessment of myocardial function on $^{201}Tl$ gated SPECT was compared with that of $^{99m}Tc$-MIBI gated SPECT Results: Correlation between the two measurements for volumes and EF was excellent by the repeated gated SPECT studies of $^{201}Tl$ (r=0.928 to 0.986; p<0.05) and $^{99m}Tc$-MIBI (r=0.979 to 0.997; p<0.05). However, Bland Altman analysis revealed the 95% limits of agreement (2 SD) for volumes and EF were tighter by repeated $^{99m}Tc$-MIBI gated SPECT (EDV: 14.1 ml, ESV: 9.4 ml and EF: 5.5%) than by repeated $^{201}Tl$ gated SPECT (EDV: 24.1 ml, ESV: 18.6 ml and EF: 10.3%). The root mean square (RMS) values of the coefficient of variation (CV) for volumes und EFs were smaller by repeated $^{99m}Tc$-MIBI gated SPECT (EDV: 2.1 ml, ESV 2.7 ml and EF: 2.3%) than by repeated $^{201}Tl$ gated SPECT (EDV: 3.2 ml, ESV: 3.5 ml and EF: 5.2%). Conclusion: $^{99m}Tc$-MIBI provides more reproducible volumes and EF than $^{201}Tl$ on repeated acquisition gated SPECT. $^{99m}Tc$-MIBI gated SPECT is the preferable method for the clinical monitoring of myocardial function.