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Changes in thoracolumbar fascial thickness, structure and shear strain are associated with lower back pain (LBP). Therapeutic taping techniques such as Kinesio-Taping (KT) are increasingly used to treat LBP, albeit with variable effects and unclear mechanisms. However, evidence for quantifying how treatment effects in vivo fascia properties is inadequate. We therefore aimed to explore taping mechanisms using an in vivo ultrasound measurement.Thoracolumbar ultrasound videos of known orientations and positions were taken from 12 asymptomatic participants (8 males and 4 females, aged 22.9 ± 3.59) while performing velocity-guided lumbar flexion with and without KT applied. An automated algorithm using cross-correlation to track contiguous tissue layers across sequential frames in the sagittal plane, was developed and applied to two movements of each subject in each taping condition. Differences of inter-tissue movements and paracutaneous translation at tissue boundaries were compared.Significant reduction in the mean movement of subcutaneous tissue during lumbar flexion before and after taping was found. There was no difference in other observed tissue layers. Tissue paracutaneous translations at three boundaries were significantly reduced during lumbar flexion when KT was applied (skin-subcutaneous: 0.25 mm, p 0.01; subcutaneous-perimuscular tissue: 0.5 mm, p = 0.02; and perimuscular-muscle: 0.46, p = 0.05). No overall reduction in lumbar flexion was found (p = 0.10).KT reduced subcutaneous inter-tissue movement and paracutaneous translation in the superficial thoracolumbar fascia during lumbar flexion, and the relationship of such difference to symptomatic change merits exploration. Combining ultrasound data with muscle activation information may be useful to reveal potential mechanisms of therapeutic taping in patients with LBP.