Torsional properties of single fibers have considerable effects on the performance of fabrics. A new torsion tester with high resolution is developed based on the torsion-balance method and the autocollimation principle for measuring the torsional properties of single fibers. The tester can measure the torque accurately using the torsion wire of known torsional rigidity. The angular displacement of the torsion wire is measured by a non-contact electronic autocollimator. The calibration results show that the maximum torque capability is around 6 × 10−6 N m with a theoretical resolution of about 8 × 10−11 N m. The instrument’s validity is verified by comparing the measurement result with that obtained by a torsion pendulum. Torsion tests on four industrially microfibers, including glass fiber, polyimide fiber, carbon fiber, and gold wire, are performed with the tester, which gives shear moduli 71.0 ± 0.4, 5.6 ± 0.1, 22.6 ± 3.1, and 34.3 ± 0.5 GPa, respectively. Young’s moduli of these fibers are also provided. It is demonstrated that these fibers have a high anisotropy in mechanical properties.