A large gap pin‐to‐plate, atmospheric‐pressure plasma reactor is demonstrated as means of in vitro study of plasma species interactions with cell cultures. By employing optical emission and optical absorption spectroscopy, we report that the pin‐to‐pate plasma array had an optimal discharge frequency for cell death of 1000 Hz in ambient air for the target cancer cell line, human glioblastoma multiform (U‐251MG). The detected plasma chemistry contained reactive oxygen and nitrogen species including OH, N2, N2+ and O3. We show that by varying the plasma discharge frequency, the plasma chemistry can be tailored to contain up to 8.85 times higher levels of reactive oxygen species (ROS) as well as a factor increase of up to 2.86 for levels of reactive nitrogen species (RNS). At higher frequencies, ROS are more dominant than RNS, which allows for a more dynamic and controlled environment for sample study without modifying the inducer gas conditions. When used for treatment of culture media and cell cultures, variation of the plasma discharge frequency over the range 1000–2500 Hz demonstrated a clear dependence of the responses, with the highest cytotoxic responses observed for 1000 Hz. We propose that the reactor offers a means of studying plasma–cell interactions and possible cofactors such as pro‐drugs and nanoparticles for a large volume of samples and conditions due to the use of well plates.

Glioblastoma multiforme (GBM) is the most common and biologically aggressive brain tumour. The current standard therapy for GBM consists in surgical resection, followed by radiotherapy and chemotherapy. Yet, the treatment is limited due to the area for the surgical resection and for the inability of some drugs to cross the brain blood barrier, leading to a general prognostic of no more than a year. Cold atmospheric plasma (CAP) is a new approach in the treatment of this challenging disease. CAP interaction with cells is dependent on physical and chemical factors, with different plasma discharges, cell type, and culture conditions leading to different CAP activity. Considering the plasma self-adaptation that different plasma discharge modes can undergo, which leads to different interaction plasma/cells, the characterization of a new device is essential. In this study we analysed the effect of a novel large pin-to-plate non-thermal atmospheric plasma on U-251 MG cells under different conditions. The analysis of reactive oxygen and nitrogen species (RONS) on plasma, media and cells were also assessed. We were able to demonstrate that the pin-to-plate device is cytotoxic to GBM cells in a dose, time and ROS dependent manner. The measurements of RONS on plasma/media also give us an insight on the chemical effect of this novelty device, and the possibility to better understand the use of this device as a promising GBM therapy.