Refining in papermaking plays an important role in changing fiber properties as well as paper properties. The major effects of refining on pulp fibers are internal and external fibrillation, fiber shortening, and fines formation. Many workers showed that internal fibrillation of the primary refining effects was most influential in improving paper properties. In particular, refining produces separation of fiber walls into several lamellae, thus causing fiber wall swelling with water penetration. This leads to the increase of fiber flexibility and of fiber-to-fiber contact during drying. If the fibers are very flexible, they will be drawn into close contact with each other by the force of surface tension as the water is removed during the drainage process and drying stages. In order to study the effect of fiber wall delamination on paper properties, cross-sectional image of fibers in a natural condition had to be generated without distortion. Finally, it was well recognized that confocal laser scanning microscope (CLSM) could be one of the most efficient tool for creating and quantifying fiber wall delamination in combination with image analysis technique. In this study, the CLSM could be used not only to observe morphological features of transverse views of swollen fibers refined under low and high intensity, but also to investigate the sequence of fiber wall delamination and fiber wall breakage. From the CLSM images, increasing the specific energy or refining decreased the degree of fiber collapse, fiber cross-sectional area, fiber wall thickness and lumen area. High intensity refining produced more external fibrillation.