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As interest grows in using tumor organoids as models for drug discovery and precision medicine, platforms for the reliable functional testing and analysis of these organoids are needed. Existing functional approaches in precision medicine face difficulties in the creation, scaling, and analysis of physiological tumor organoid models. We present a method for the automated bioprinting of tumor organoids coupled with real-time biomass quantification using high-speed live cell interferometry (HSLCI). This workflow enhances consistency, preserves sample viability, and assesses response to treatment with single-organoid resolution. First, we describe the bioprinting protocol used to achieve the deposition of organoids in uniformly thin three-dimensional layers of Matrigel. Then we show that bioprinting preserves the histological and molecular characteristics of manually seeded organoids. In drug screening experiments, we demonstrate the simultaneous monitoring of thousands of individual organoids and show that HSLCI can rapidly identify drug sensitivity and resistance to treatment. We observe significant heterogeneity in the drug responses of single organoids and discuss the potential implications for use of this pipeline in the clinical setting. Citation Format: Peyton J. Tebon, Bowen Wang, Alexander L. Markowitz, Graeme Murray, Ardalan Davarifar, Huyen T. Nguyen, Nasrin Tavanaie, Thang L. Nguyen, Paul C. Boutros, Michael A. Teitell, Alice Soragni. High-speed live cell interferometry captures heterogeneity in bioprinted tumor organoids [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 3081.