Quantity and quality assessment of human pancreatic islets are essential processes to define a safe and potent quality product used for clinical transplantation. The conventional method of manual assessment has been used in the field for longer than two decades. The high degree of variability in product quantity and lack of archival imaging records of the product for verification are two major disadvantages of using the manual method for quantity and quality assessment of human pancreatic islets. Investigators have developed promising new methods for technical improvement. In this study, we briefly review the published methods and highlight the advantages of digital imaging analysis (DIA) when compared to the manual method. The application of DIA reduces measurement variability and increases the precision of islet equivalent (IEQ) determination for batch analysis. It produces images that can be archived for retrospective analysis and validation, and the data can be transmitted electronically for off-site analysis. These features are important for quality pancreatic islet assessment and are consistent with FDA requirements of current good manufacturing practice for clinical islet transplantation.