The quantification of knee alignment is a routine part of orthopaedic practice and is important for monitoring disease progression, planning interventional strategies, and follow-up of patients. Currently available technologies such as radiographic measurements have a number of drawbacks. The aim of this study was to validate a potentially improved technique for measuring knee alignment under different conditions. An image-free navigation system was adapted for non-invasive use through the development of external infrared tracker mountings. Stability was assessed by comparing the variance (F-test) of repeated mechanical femoro-tibial (MFT) angle measurements for a volunteer and a leg model. MFT angles were then measured supine, standing and with varus-valgus stress in asymptomatic volunteers who each underwent two separate registrations and repeated measurements for each condition. The mean difference and 95% limits of agreement were used to assess intra-registration and inter-registration repeatability. For multiple registrations the range of measurements for the external mountings was 1 larger than for the rigid model with statistically similar variance ( p ¼ 0.34). Thirty volunteers were assessed (19 males, 11 females) with a mean age of 41 years (range: 20-65) and a mean BMI of 26 (range: 19-34). For intra-registration repeatability, consecutive coronal alignment readings agreed to almost AE1, with up to AE0.5 loss of repeatability for coronal alignment measured before and after stress maneuvers, and a AE0.2 loss following stance trials. Sagittal alignment measurements were less repeatable overall by an approximate factor of two. Inter-registration agreement limits for coronal and sagittal supine MFT angles were AE1.6 and AE2.3 , respectively. Varus and valgus stress measurements agreed to within AE1.3 and AE1.1 , respectively. Agreement limits for standing MFT angles were AE2.9(coronal) and AE5.0 (sagittal), which may have reflected a variation in stance between measurements. The system provided repeatable, real-time measurements of coronal and sagittal knee alignment under a number of dynamic, real-time conditions, offering a potential alternative to radiographs.