Purpose <p>Synaptic vesicle glycoprotein 2A (SV2A) is a critical biomarker for evaluating synaptic density in neurological research. Among available radioligands, [¹⁸F]SynVesT-1 is increasingly used in PET research because of its extended half-life, while having comparable pharmacokinetic properties to the widely used [¹¹C]UCB-J. However, quantitative application in rat models remains unexplored for [¹⁸F]SynVesT-1. This study aims to validate quantitative kinetic modelling methods for [¹⁸F]SynVesT-1 and develop non-invasive quantification methods for synaptic density in rats.</p> Methods <p>First, blood analysis of [¹⁸F]SynVesT-1 was performed to generate metabolite-corrected plasma input functions. Then, kinetic modelling was evaluated using compartmental analysis approaches, as well as Logan plot. Furthermore, non-invasive image-derived input functions (IDIF), with and without non-negative matrix factorization (NMF) were compared against the arterial input function (AIF).</p> Results <p>Blood analysis showed that the parent fraction of the tracer decreased over time following a sigmoid curve, while the plasma-to-whole blood ratio remained stable over time (0.89 ± 0.02). The two-tissue compartmental model (2TCM) and Logan plot were determined to be the most accurate methods for quantification of [¹⁸F]SynVesT-1 kinetics in rats. Additionally, the results demonstrated strong agreement between AIF-derived and image-derived volume of distribution (<i>V</i>_T) values, with both image-derived input approaches (IDIF and IDIF-NMF) performing equally well.</p> Conclusion <p>These findings validate kinetic modelling methods for [¹⁸F]SynVesT-1 PET, enabling their application in further rat studies for preclinical neuroscience research and prove that image-derived input functions are reliable non-invasive alternatives to AIF.</p>