The cross-section area of animal fibers varies along the fiber length, and this geometrical irregularity has a major impact on the mechanical properties of those fibers. In practice fibers are often subjected to tensile stresses during processing and application, which may change fiber cross-section area. It is thus necessary to examine geometrical irregularity of fibers under tension. In this study, scoured animal fibers were subjected to different tensile loading using a Single Fiber Analyzer (SIFAN) instrument. The 3D images of the fiber specimens were first constructed, and then along-fiber diameter irregularities of the specimens were analyzed for different levels of tensile loading. The changes in effective fineness of the fiber specimens were also discussed. The results indicate that for the wool fibers examined, there is considerable discrepancy in the fiber diameter results obtained from the commonly used single scan along fiber length and that from multiple scans at different rotational angles, and that the diameter variation along fiber length increases as fiber tension increases. The results also show that when diameter reduction treatments are applied to wool by stretching, the reduced average fiber diameter is associated with an increase in both within-fiber and between-fiber diameter variations. So in terms of effective fineness, the change is much smaller than the difference between the average diameters of the parent and treated wool. These results have significant implications for improving the accuracy of fiber diameter measurement and evaluation.