The effect of microstructural modifications of V2O5 thin films, obtained through alterations in post oxidation duration, on methane sensing behavior is reported for the first time. Three different oxidation times viz., 1 h, 3 h and 5 h yielded varied microstructure and vibrational properties as evident from XRD and Raman investigation. These changes in properties manifest as differences in gas sensing behavior. Methane sensing properties of V2O5 was investigated in temperature range from 100 to 300 °C and optimum operating temperature of 200 °C was identified for all three samples. Films oxidized for 1 h showed the highest response due to favorable surface conditions which are discussed. These results will help in tailoring microstructure towards device level application processes.