Purpose: The purpose of this study was to determine the lowest Tl-201 dose that does not reduce the image quality of myocardial perfusion SPECT (MPS) by Poisson resampling simulation. Methods: One hundred and twelve consecutive MPS data from patients with suspected or known coronary artery disease were collected retrospectively. Stress and rest MPS data were resampled using the Poisson method with 33%, 50%, 67%, and 100% count settings. Two nuclear medicine physicians assessed the image quality of reconstructed data visually by giving grades from − 2 to + 2. The summed stress score (SSS), summed rest score (SRS), and summed difference score (SDS) were obtained on the workstation. Image quality grades and semi-quantitative scores were then compared among these resampled images. Results: The proportions of “adequate” image quality were 0.48, 0.75, 0.92, and 0.96 for the groups of images with 33%, 50%, 67%, and 100% data, respectively. The quality of the resampled images was significantly degraded at 50% and 33% count settings, while the image quality was not different between 67 and 100% count settings. We also found that high body mass index further decreased image quality at 33% count setting. Among the semi-quantitative parameters, SSS and SRS showed a tendency to increase with a decline in count. Conclusion: Based on the simulation results, Tl-201 dose for MPS can be reduced to 74 MBq without significant loss of image quality. However, the SSS and SRS can be changed significantly, and it needs to be further verified under the different conditions.