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The combination of organoid culture techniques with mass spectrometry imaging provides a label\u2010free methodology for characterizing drug penetration, patient\u2010specific response, and drug biotransformation. However, current methods used to grow tumor organoids employ extracellular matrices that can produce small molecule background signal during mass spectrometry imaging analysis. Here, we develop a method to isolate 3D human tumor organoids out of a Matrigel extracellular matrix into gelatin mass spectrometry compatible microarrays for high\u2010throughput mass spectrometry imaging analysis. The alignment of multiple organoids in the same z\u2010axis is essential for sectioning organoids together and for maintaining reproducible sample preparation on a single glass slide for up to hundreds of organoids. This method successfully removes organoids from extracellular matrix interference and provides an organized array for high\u2010throughput imaging analysis to easily identify organoids by eye for area selection and further analysis. With this method, mass spectrometry imaging can be readily applied to organoid systems for preclinical drug development and personalized medicine research initiatives.<\/jats:p>","DOI":"10.1002\/jms.4452","type":"journal-article","created":{"date-parts":[[2019,10,29]],"date-time":"2019-10-29T19:39:48Z","timestamp":1572377988000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":34,"title":["Sample preparation strategies for high\u2010throughput mass spectrometry imaging of primary tumor organoids"],"prefix":"10.1002","volume":"55","author":[{"given":"Jillian","family":"Johnson","sequence":"first","affiliation":[{"name":"School of Pharmacy University of Wisconsin\u2014Madison  Madison Wisconsin"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Joe T.","family":"Sharick","sequence":"additional","affiliation":[{"name":"Morgridge Institute for Research  Madison 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