Abstract The goal of personalized oncology is to select the most appropriate drug(s) for individual cancers. A number of platforms (e.g., organoids and PDX mice) have been developed to generate human cancer models for drug sensitivity testing, but they have yet to deliver on their promise as an every-day clinical assay for use by oncologists. The major hurdles limiting success of personalized drug sensitivity testing are the long turnaround time and the amount of starting material needed, in addition to the loss of the original tumor microenvironment. To address these shortcomings, we developed an elegant assay to evaluate drug sensitivity based on metabolic changes in fresh tumor tissues obtained through needle biopsies. Our so-called MetaboCore assay takes advantage of an optimized organotypic culture platform that is suitable drug testing immediately upon specimen collection, and a novel single-cell metabolic assay that detects changes in metabolism of cancer cells within a short time of drug exposure. Consequently, quantitative results of relative drug sensitivity can be obtained within a week of the biopsy. In order to advance our assay towards clinical use, we need to create a robust SOP with defined parameters for each step of the assay in order to achieve reproducible results. The objectives of the proposal are to examine how biospecimen preanalytical conditions affect assay performance. Specifically, we will quantify the effects of each of the following variables on tissue viability and drug response: type and size of biopsy needles, transport solution, overnight storage, culture conditions, recovery time, drug concentration, treatment duration, and protocols for tissue dissociation. Based on these findings, we will create a SOP for MetaboCore and test its clinical performance in a pilot study using needle biopsies of human liver cancers.