We investigated the in vivo length changes of the collateral ligaments of metacarpophalangeal joint during flexion. We obtained computed tomography scans of index, middle, ring and little fingers at 0°, 30°, 60° and 90° of joint flexion from six hands of six healthy adult volunteers. Three of them had their dominant right hand studied, and the other three had their non-dominant left hand studied. We measured and analysed the radial and ulnar collateral ligaments of each metacarpophalangeal joint from the reconstructed images. We found that the dorsal and middle portions of the both radial and ulnar collateral ligament lengthened progressively during digital flexion and reached the maximum at 90° flexion. The length of the volar portion increased from 0° to 30° flexion and then decreased from 30° to 60° flexion, reaching the minimum at 90°. In conclusion, three portions of collateral ligaments on both sides of the metacarpophalangeal joint have variable length changes during flexion, which act to stabilize the joint through its flexion arc.