Objective To quantitatively evaluate the effect of 2 types of pressures induced injury by using threedimensional (3D) reconstruction of rats loaded tibial is anterior muscle from two-dimensional (2D) image of serial histological sections. Methods Twenty female or male Sprague Dawley rats, aged 10-12 weeks and weighing 280-300 g, were randomlydivided into experimental group (n=10) and control group (n=10). The random side of tibial is anterior muscle was givenintermittent gradient (8.0-21.3 kPa) and sustained (13.3 kPa) pressure in 0.12 cm2 area in experimental group and controlgroup, respectively; the experiment was terminated and the general condition of rats was observed after 3 cycles, and a single cycle included 2 hours of compression and 30 minutes of release. The general observations of pressed skin and tibial is anterior muscle were done after 24 hours of pressure rel ief, and the tibial is anterior muscle was harvested integrally from the loaded side, then made into interval 4 μm serial sections. After HE staining, 2D images were obtained. Necrosis and injury areas were distinguished by Image Pro Plus (IPP) 6.0 software and image registration was conducted by Photoshop 8.0.1 after 2D panorama images acquired by digital microscope (× 40) and IPP mosaic software. 3D reconstruction was establ ished via data processing using Mimics 10.1 software so as to get the volume, the surface area, and 3D images of the whole piece of tibial is anterior muscle and injury areas respectively. Results All rats of 2 groups survived till experiment terminated and no skin ulcers occurred after 24 hours. Edema and indentation were observed on press side skin and tibial is anterior muscles of 2 groups, fadeless maroon area was observed in control group. A total of 994 sl ices were obtained from 20 samples of tibial is anterior muscles. 3D images suggested that injury of control group was severe, which penetrated the whole piece of tibial is anterior muscle and expandedalong the tibia bony prominence. By contrast, injury of experimental group was less, but had similar width to the contact surface of indentor. There was no significant difference in the volume and the surface area of tibial is anterior muscle between 2 groups (P gt; 0.05), while the injury volume and the injury surface area were significantly smaller in experimental group than in control group (P lt; 0.05). Conclusion 3D reconstruction is an effective method to quantitatively evaluate pathological changes inside the integrity tissue and can provide the visual basis for the mechanical property distributed in the loaded muscle. Intermittent gradient pressure can reduce deep tissue injury.