Clinical and experimental studies have shown that several mechanical factors influence the fracture healing process. One such factor, interfragmentary movement, is affected by loading and the stiffness of the fixation device. This study evaluated the stiffness of different external fixation devices for a rat femoral fracture model, using in vitro and analytical methods. The contribution to the stiffness of the fixation construct was dominated by the flexibility of the pins in relation to their offset, diameter, and material properties. The axial stiffness increased with decreasing offset and increasing pin diameter. Titanium pins resulted in significantly lower axial stiffness compared to stainless steel pins of the same design. The fixator body material and fixator length had a less pronounced influence on fixation stiffness. Mechanically characterized external fixation devices will allow in vivo study of the fracture healing process utilizing pre‐calculated fracture fixation stiffness. These characterized fixation devices will allow controlled manipulation of the axial and shear interfragmentary movement to achieve a flexible fixation resulting in callus formation compared to a more rigid fixation limiting callus formation in a rat femoral fracture model. © 2008 Orthopaedic Research Society. Published by Wiley Periodicals, Inc. J Orthop Res 27: 687–693, 2009