FeCrAl-based materials have attracted ever-increasing attentions due to their excellent resistances against high-temperature oxidation and stress-corrosion cracking, while enhancement of their mechanical properties can expand their potential applications. In this work, the micro-alloying method by adding Nb was applied to enhance the mechanical property of FeCrAl. The FeCrAl thin films with different Nb contents (Fe-16Cr-5Al, Fe-14Cr-5Al-1Nb, Fe-12Cr-5Al-2Nb, Fe-13Cr-3Al-7Nb, in wt%) were synthesized using magnetron co-sputter deposition. Benefiting from the refined grain size, the as-deposited Fe-13Cr-3Al-7Nb thin film exhibited the highest hardness (3.3 GPa), followed by Fe-14Cr-5Al-1Nb (1.5 GPa) and Fe-12Cr-5Al-2Nb (1.2 GPa), whereas the hardness of Nb-free film was the lowest (0.8 GPa). The fracture resistance (which is related to the ratio of hardness and elastic modulus) also enhances with the addition of Nb. After annealing (873 K for 6 h), the hardness of the Fe-13Cr-3Al-7Nb thin film further increased to ~6.7 GPa, in contrast to the as-annealed Nb-free thin film with no obvious hardness change. The CALPHAD (CALculation of PHAse Diagram) results indicate that with adding Nb the original single BCC (body centered cubic) phase tended to decompose into the Fe-rich and Cr-rich BCC domains, which is consistent with the transmission electron microscopic (TEM) observations. The observed strengthening of the annealed Nb-containing samples is therefore attributed to the phase separation. The first-principles calculations also support this notion. Thus, the current work has delineated the effect of Nb addition on both deposited and annealed FeCrAl-based thin films, which shows the usefulness of combined experimental and computational methods in interpreting microstructure and mechanical property evolutions of engineering materials. Display omitted •Grain refinement strengthens the as-deposited FeCrAl thin films by adding Nb.•Phase separation strengthens the as-annealed FeCrAl thin films by adding Nb.•The microstructure and mechanical property evolutions were well interpreted by experimental and computational methods.