Pulsed light (PL) is a non-thermal technology which leads to different applications (food, pharmaceutical and cosmetic packaging, medical devices) by inactivating microorganisms, knowing that the properties of PL technology are cumulative treatment, especially for bacteria. PL uses intense flashes of white light (200 nm–1100 nm wavelengths), rich in UV (200 nm–400 nm) and produced by xenon flash lamps. Localized photothermal and photophysical effects, but also the capability of its UV light component to modify the structure of biomolecules (photochemical effect), such as DNA and proteins, are the main mechanisms involved to explain the inactivation of microorganisms by PL. As compared to other sterilization methods, such as heat or chemical disinfectant treatment, PL treatment has several advantages; it is faster, induces low operational cost and leaves no residues. Nevertheless, no exhaustive data on exposure conditions are available, which would make it possible to repeat, interpret and evaluate the data needed for large-scale industrial deployment of PL in pharmaceutical and cosmetic packaging and medical devices. Research should also focus on understanding the inactivation mechanisms of different bacterial strains, states, and morphologies when exposed to PL.