en Cellular and molecular Neuroscience Cellular and molecular Neuroscience Original Original <div style="text-align: justify;"><strong>Introduction: </strong>Organotypic Hippocampal Brain Slices (OHBS) provide an advantageous alternative to in vivo models to scrutinize Traumatic Brain Injury (TBI). We followed a well-established TBI protocol, but noticed that several factors may influence the results in such a set-up. Here, we describe a structured approach to generate more comparable results and discuss why specific eligibility criteria should be applied.<br> <strong>Methods: </strong>We defined necessary checkpoints and developed inclusion and exclusion criteria that take the observed variation in such a model into consideration. Objective measures include the identification and exclusion of pre-damaged slices and outliers. Six steps were outlined in this study.<br> <strong>Results: </strong>A six-step approach to enhance comparability is proposed and summarized in a flowchart. We applied the suggested measures to data derived from our TBI-experiments examining the impact of three different interventions in 1459 OHBS. Our exemplary results show that through equal requirements set for all slices more precise findings are ensured.<br> <strong>Conclusion:</strong> Results in a TBI experiment on OHBS should be analyzed critically as inhomogeneities may occur. In order to ensure more precise findings, a structured approach of comparing the results should be followed. Further research is recommended to confirm and further develop this framework.</div> Organotypic hippocampal brain slices, In vitro model, Traumatic Brain Injury (TBI), Propidium Iodide (PI), Eligibility, Framework 485 498 http://bcn.iums.ac.ir/browse.php?a_code=A-10-1213-1&slc_lang=en&sid=1 Linda Grüßer linda.gruesser@googlemail.com 13700319475328460030846 13700319475328460030846 Yes Department of Anesthesiology, RWTH Aachen University Hospital, Aachen, Germany. Rosmarie Blaumeiser-Debarry Rblaumeiser-debarry@ukaachen.de 13700319475328460030847 13700319475328460030847 No Department of Anesthesiology, RWTH Aachen University Hospital, Aachen, Germany. Rolf Rossaint rrossaint@ukaachen.de 13700319475328460030848 13700319475328460030848 No Department of Anesthesiology, RWTH Aachen University Hospital, Aachen, Germany. Matthias Krings matthias.krings@rwth-aachen.de 13700319475328460030849 13700319475328460030849 No Department of Anesthesiology, RWTH Aachen University Hospital, Aachen, Germany. Benedikt Kremer benedikt.kremer@rwth-aachen.de 13700319475328460030850 13700319475328460030850 No Department of Neurosurgery, RWTH Aachen University Hospital, Pauwelsstr. 30, 52074 Aachen, Germany Anke Höllig ahoellig@ukaachen.de 13700319475328460030851 13700319475328460030851 No Department of Neurosurgery, RWTH Aachen University Hospital, Aachen, Germany. Mark Coburn mcoburn@ukaachen.de 13700319475328460030852 13700319475328460030852 No Department of Anesthesiology, RWTH Aachen University Hospital, Aachen, Germany.