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Purpose: Using microdosimetry, this study investigated the relative biological effectiveness (RBE) and quality factor (Q¯) variations in field and out of field as a function of radiation quality for clinical protons., Methods and Materials: A water phantom with a spread-out Bragg peak (SOBP) was irradiated to acquire microdosimetric spectra at several distal and lateral depths with a tissue equivalent proportional counter. The measurements were used as inputs to microdosimetric kinetic and Loncol models to determine the RBE spatial distribution and compare it with predictions from the dose-averaged linear energy transfer-based McNamara model. Q¯ values and biological and dose equivalent values were also calculated., Results: The data demonstrated that radiation quality changed more rapidly with depth than lateral distance from the SOBP. In beam, y D ranged from approximately 4 keV/μm at the entrance to 8 keV/μm at the SOBP far end, reaching approximately 15 keV/μm at the penumbra. Out of field, the overall highest value of 23 ± 2 keV/μm was observed at the beam-edge penumbra. Radiation quality changes caused RBE deviations from the clinical value of 1.1, whose extent depends on the approach used for assessing radiation quality as well as on the radiobiological model. For RBE 10 , microdosimetry-based models appeared to better reproduce the radiobiological data than the dose-averaged linear energy transfer model. Out of field, both the RBE and Q¯ values appeared to have limitations in describing the radiation biological effectiveness. This research also presents a first comprehensive benchmark of TOPAS code against in-field and out-of-field microdosimetric spectra of therapeutic protons., Conclusions: Further investigation will be necessary to evaluate the quantitative effects of RBE variations on treatment planning and assess the clinical consequences in terms of both tumor control and normal-tissue toxicity. The achievement of this goal calls for accurate radiobiological data to validate the RBE models., (Copyright © 2022 Elsevier Inc. All rights reserved.)