Quantum wells and barriers with precise thicknesses and abrupt composition changes at their interfaces are critical for obtaining the desired emission wavelength from quantum cascade laser devices. High-resolution X-ray diffraction and transmission electron microscopy are commonly used to calibrate and characterize the layers’ thicknesses and compositions. A complementary technique, atom probe tomography, was employed here to obtain a direct measurement of the 3-dimensional spatially-resolved compositional profile in two InxGa1−xAs/InyAl1−yAs III-V strained-layer superlattice structures, both grown at 605 °C. Fitting the measured composition profiles to solutions to Fick’s Second Law yielded an average interdiffusion coefficient of 3.5 × 10−23 m2 s−1 at 605 °C. The extent of interdiffusion into each layer determined for these specific superlattices was 0.55 nm on average. The results suggest that quaternary active layers will form, rather than the intended ternary compounds, in structures with thicknesses and growth protocols that are typically designed for quantum cascade laser devices.