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A pencil beam proton radiography (PR) method, using a commercial multilayer ionization chamber (MLIC) integrated with a treatment planning system (TPS) was developed. A Giraffe (IBA Dosimetry) MLIC (±0.5 mm accuracy) was used to obtain pencil beam PR by delivering spots uniformly positioned at a 5.0 mm distance in a 9  ×  9 square of spots. PRs of an electron-density (with tissue-equivalent inserts) phantom and a head phantom were acquired. The integral depth dose (IDD) curves of the delivered spots were computed by the TPS in a volume of water simulating the MLIC, and virtually added to the CT at the exit side of the phantoms. For each spot, measured and calculated IDD were overlapped in order to compute a map of range errors. On the head-phantom, the maximum dose from PR acquisition was estimated. Additionally, on the head phantom the impact on the range errors map was estimated in case of a 1 mm position misalignment. In the electron-density phantom, range errors were within 1 mm in the soft-tissue rods, but greater in the dense-rod. In the head-phantom the range errors were  −0.9  ±  2.7 mm on the whole map and within 1 mm in the brain area. On both phantoms greater errors were observed at inhomogeneity interfaces, due to sensitivity to small misalignment, and inaccurate TPS dose computation. The effect of the 1 mm misalignment was clearly visible on the range error map and produced an increased spread of range errors (−1.0  ±  3.8 mm on the whole map). The dose to the patient for such PR acquisitions would be acceptable as the maximum dose to the head phantom was  <2cGyE. By the described 2D method, allowing to discriminate misalignments, range verification can be performed in selected areas to implement an in vivo quality assurance program. [ABSTRACT FROM AUTHOR]