Objective. Image reconstruction in high resolution, narrow bore PET scanners with depth of interaction (DOI) capability presents a substantial computational challenge due to the very high sampling in detector and image space. The aim of this study is to evaluate the use of a virtual cylinder in reducing the number of lines of response (LOR) for DOI-based reconstruction in high resolution PET systems while maintaining uniform sub-millimetre spatial resolution. Approach. Virtual geometry was investigated using the awake animal mousePET as a high resolution test case. Using GEANT4 Application for Tomographic Emission (GATE), we simulated the physical scanner and three virtual cylinder implementations with detector size 0.74 mm, 0.47 mm and 0.36 mm (vPET1, vPET2 and vPET3, respectively). The virtual cylinder condenses physical LORs stemming from various crystal pairs and DOI combinations, and which intersect a single virtual detector pair, into a single virtual LOR. Quantitative comparisons of the point spread function (PSF) at various positions within the field of view (FOV) were compared for reconstructions based on the vPET implementations and the physical scanner. We also assessed the impact of the anisotropic PSFs by reconstructing images of a micro Derenzo phantom. Main results. All virtual cylinder implementations achieved LOR data compression of at least 50% for DOI PET reconstruction. PSF anisotropy in radial and tangential profiles was chiefly influenced by DOI resolution and only marginally by virtual detector size. Spatial degradation introduced by virtual cylinders was most prominent in the axial profile. All virtual cylinders achieved sub-millimetre volumetric resolution across the FOV when 6-bin DOI reconstructions (3.3 mm DOI resolution) were performed. Using vPET2 with 6 DOI bins yielded nearly identical reconstructions to the non-virtual case in the transaxial plane, with an LOR compression factor of 86%. Resolution modelling significantly reduced the effects of the asymmetric PSF arising from the non-cylindrical geometry of mousePET. Significance. Narrow bore and high resolution PET scanners require detectors with DOI capability, leading to computationally demanding reconstructions due to the large number of LORs. In this study, we show that DOI PET reconstruction with 50%–86% LOR compression is possible using virtual cylinders while maintaining sub-millimetre spatial resolution throughout the FOV. The methodology and analysis can be extended to other scanners with DOI capability intended for high resolution PET imaging.