Calibration of MTT assay in proton beams using radiochromic films.

PURPOSE: This study provides methodology of calibrating as well as controlling the output for an MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) colorimetric assay irradiated in a low energy proton beam using EBT3-model GAFCHROMICTM film, without correcting for quenching effect. METHODS: A calibrated Markus ionization chamber was used to measure the depth dose and beam output for 26.5 MeV protons produced by a CS30 cyclotron. A time-controlled aluminum cylinder was added in front of the horizontal beam-exit serving as a radiation shutter. Following the TRS-398 reference dosimetry protocol for proton beams, the output was calibrated in water at a reference depth of 3 mm. EBT3 film was calibrated for doses up to 8 Gy at the same depth. To verify the dose distribution for each 96-well MTT assay plate, EBT3 film was placed at the reference depth during irradiation and cell doses were scaled by measured percent depth dose (PDD) data. RESULTS: The radiochromic film dosimetry system in this study provides dose measurements with an uncertainty better than 3.3% for doses higher than 1 Gy. From a single exposure and utilizing the Gaussian shape of the beam, multiple dose points can be obtained within different wells of the same plate ranging from 6.9 Gy (sigma ∼4%) in the central well, and 2 Gy (sigma ∼8%) for wells positioned closer to the periphery. CONCLUSIONS: We described a methodology for radiochromic film-based dose monitoring system, using low-energy protons, which can be used for the MTT assay in any proton beam, except within Bragg peak region.

Dosimetry of biological irradiations using radiochromic films.

Delivering accurate radiation dose to blood specimens during biological irradiations is essential in quantifying damage of ionizing radiation. To estimate dose to blood samples as accurately as possible, pieces of EBT2 model GAFCHROMIC™ film were placed within an approximately 10 mm finely ground rice layer that was used to simulate test specimens inside 40 mL plastic flasks. Irradiations of flasks were carried out using an X-RAD 320 irradiator with a beam quality of 320 kVp and a measured half value layer of 1.12 mm Cu, in air and in a full scattering setup which consisted of either rice or Solid Water™ (SW) surrounding flasks, filled to the same level at top of the flasks, together with a 5 cm thick SW slab beneath them. Outputs, per cent depth doses and beam profiles at different depths were measured and compared between setups. For the same setting, the dose delivered to the middle flask under the full scattering setup is 22% larger than with the in-air setup at the depth of the specimen and 9.2% more homogeneous across the specimen thickness of 10 mm (2.3% variation in comparison to the surface). Rice showed a fairly similar performance to SW within 1% at the same depth of 10 mm. Experimental setup based on full scattering conditions was shown to provide faster, more homogenous and fairly uniform dose delivery to biological specimens in comparison to conventionally used in-air setups.