Photon planning was done with Raystation treatment planning system (RaySearch Laboratories) based on a CT scan of the hypoxia chamber containing 96-well plates filled with water. Irradiation was performed on a Siemens Artiste (6 MV). For carbon ion delivery, Spread-out Bragg peak (SOBP) was physically optimized with the Treatment Planning for Particles (TRiP98) system, using a 30x30x30 cm3 water phantom positioned at 100 cm source-surface distance (SSD) [17 (link)]. To reduce physical uncertainties on cell survival introduced by the heterogeneous target, a detailed geometry of the utilized 96-well plates was incorporated into a FLUKA Monte Carlo simulation of the Heidelberg Ion Beam Therapy (HIT) beam-line [18 (link)]. Dose maps were generated, with dose uniformity found to be within 2% range in the SOBP region. Carbon dose levels for planned 1, 2, 4 and 6 Gy were corrected accordingly to actual prescribed 0.95, 1.9, 3.8, and 5.64 Gy.
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