In this paper, we present a novel hyperspectral system designed for imaging the human iris in vivo. The instrument is based on a tunable light source (from 480 nm to 900 nm) and a monochrome camera. The system can acquire hyperspectral cubes in 4 s, with spatial and spectral resolutions of 11 μm and 20 nm, respectively. The optical characterization of the new instrument is described, and we demonstrate its safety with respect to ANSI standards. Preliminary in vivo measurements in human volunteers are also presented.