Currently, there are no standards or guidelines for producing statistically significant data using AFIS, the Advanced Fiber Information System. For this study AFIS generates fiber measurements from 5000 individual fibers from each of 25 hand blended slivers. This research addresses key issues, such as sampling techniques, minimum sample size and test conditions and how these effect AFIS measurements. Two versions of AFIS were also compared to each other and to more traditional measurement techniques. The results of the research indicate that humidity affects maturity measurements, under controlled temperature and humidity, sampling is not as critical as for non-controlled testing conditions. In Standard Conditions, using Version 2 the minimum sample should contain at least 75,000 fibers. AFIS Version 4 has better measures of length than AFIS Version 2 due to the new nozzle straightening the fibers. Sample preparation is also very important for maturity measurements, especially IFF and Theta CV from AFIS Version 2. AFIS Version 2's individual fiber immaturity properties, Theta CV, and IFF, have the highest direct correlation to image analysis measurements of white specks. As the Circularity CV and IFF decreased, the white speck decreased. Conversely, as Micronafis increased, white speck decreased. Similarly, IFC is a good indicator of immaturity for AFIS Version 4, and as the ratio of IFC/MFC increases, white speck levels increase. Version 4's IFC and MFC provided the strongest correlation to white specks.