Potential sterilization/disinfection of medical devices (MDs) is investigated using a specific plasma process developed at the Université de Montréal over the last decade. The inactivating medium of the microorganisms is the flowing afterglow of a reduced-pressure N2-O2 discharge, which provides, as the main biocidal agent, photons over a broad ultraviolet (UV) wavelength range. The flowing afterglow is considered less damaging to MDs than the discharge itself. Working at gas pressures in the 400—700 Pa range (a few torr) ensures, through species diffusion, the uniform filling of large volume chambers with the species outflowing from the discharge, possibly allowing batch processing within them. As a rule, bacterial endospores are used as bio-indicators (BI) to validate sterilization processes. Under the present operating conditions, Bacillus atrophaeus is found to be the most resistant one and is therefore utilized as BI. The current paper reviews the main experimental results concerning the operation and characterization of this sterilizer/disinfector, updating and completing some of our previously published papers. It uses modeling results as guidelines, which are particularly useful when the corresponding experimental data are not (yet) available, hopefully leading to more insight into this plasma afterglow system. The species flowing out of the N2-O2 discharge can be divided into two groups, depending on the time elapsed after they left the discharge zone as they move toward the chamber, namely the early afterglow and the late afterglow. The early flowing afterglow from a pure N2 discharge (also called pink afterglow) is known to be comprised of N2+ and N4+ ions. In the present N2-O2 mixture discharge, NO+ ions are additionally generated, with a lifetime that extends over a longer period than that of the nitrogen molecular ions. We shall suppose that the disappearance of the NO+ ions marks the end of the early afterglow regime, thereby stressing our intent to work in an ion-free process chamber to minimize damage to MDs. Therefore, operating conditions should be set such that the sterilizer/disinfector chamber is predominantly filled by N and O atoms, possibly together with long-lived metastable-state O2(1 Δg) (singlet-delta) molecules. Various aspects related to the observed survival curves are examined: the actual existence of two “phases” in the inactivation rate, the notion of UV irradiation dose (fluence) and its implications, the UV photon best wavelength range in terms of inactivation efficiency, the influence of substrate temperature and the reduction of UV intensity through surface recombination of N and O atoms on the object/packaging being processed. To preserve their on-shelf sterility, MDs are sealed/wrapped in packaging material. Porous packaging materials utilized in conventional sterilization systems (where MDs are packaged before being subjected to sterilization) were tested and found inadequate for the N2-O2 afterglow system in contrast to a (non-porous) polyolefin polymer. Because the latter is non-porous, its corresponding pouch must be kept unsealed until the end of the process. Even though it is unsealed, but because the opening is very small the O2(1Δg) metastable-state molecules are expected to be strongly quenched by the pouch material as they try to enter it and, as a result, only N and O atoms, together with UV photons, are significantly present within it. Therefore, by examining a given process under pouch and no-pouch conditions, it is possible to determine what are the inactivating agents operating: (i) when packaged, these are predominantly UV photons, (ii) when unpackaged, O2(1Δg) molecules together with UV photons can be acting, (iii) comparing the inactivation efficiency under both packaged and unpackaged conditions allows the determination of the relative contribution of UV photons (if any) and O2(1Δg) metastable-state molecules. Such a method is applied to pyrogenic molecules and to the enzymatic activity of lysozyme proteins once exposed to the N2-O2 flowing afterglow. Finally, the activity of the infectious prion protein is shown to be reduced when exposed to the present flowing afterglow, as demonstrated by both in vitro and in vivo experiments.