Patient radiation dose and image quality are primary issues in the conduct of nuclear medicine (NM) procedures. A range of protocols are currently used in image acquisition and analysis of quality control (QC) tests, with National Electrical Manufacturers Association (NEMA) methods and protocols widely accepted in providing an accurate description, measurement and report of γ-camera performance parameters. However, no standard software is available for image analysis. Present study compares vendor QC software analysis and three types of software freely downloadable from the internet: NMQC, NM Toolkit and ImageJ-NM Toolkit software. These were used for image analysis of QC tests of γ-cameras based on NEMA protocols including non-uniformity evaluation. Ten non-uniformity QC images were obtained using a dual head γ-camera installed in Trieste General Hospital and then analyzed. Excel analysis was used as the baseline calculation for the non-uniformity test according to NEMA procedures. The results of non-uniformity analysis showed good agreement between the independent types of software and Excel calculations (the average differences were 0.3%, 2.9%, 1.3% and 1.6% for the Useful Field of View (UFOV) integral, UFOV differential, Central Field of View (CFOV) integral and CFOV differential, respectively), while significant differences were detected following analysis using the company QC software when compared with Excel analysis (the average differences were 14.6%, 20.7%, 25.7% and 31.9% for the UFOV integral, UFOV differential, CFOV integral and CFOV differential, respectively). Compared to use of Excel calculations use of NMQC software was found to be in close accord. Variation in results obtained using the three types of software and γ-camera QC software was due to the use of different pixel sizes. It is important to conduct independent analyses tests in addition to using the vendor QC software in order to determine the differences between values.