Background: Measurement of microfibril angle (MFA) in the S2 layer of the cell walls of wood using speed of sound in wood has become a common way to estimate wood stiffness and stability. Measurements across the radius of a disc can be critical, but sonic testing needs to be validated against direct measurements of MFA. Methods: A robotic machine that uses two transducers to detect the speed of sound through discs of wood from tree stems was validated by comparing its measurements with wood MFA, air-dry density, and modulus of elasticity assessed by X-ray diffractometry through the "Silviscan" system. Results: Ultrasonic velocity (USV) was highly correlated with MFA but not with air-dry density. Variable moving averages were assessed in order to estimate the resolution of the USV measurements, and it appeared that the diameter of the swath detected by the transducers was between 23 and 30 mm. Conclusions: It was concluded that MFA in wood samples can be assessed rapidly and cheaply using USV measurements but at lower resolution than assessments using X-ray diffractometry.