TY - JOUR T1 - Multiplexed and reproducible high content screening of live and fixed cells using the Dye Drop method JF - bioRxiv DO - 10.1101/2021.08.27.457854 SP - 2021.08.27.457854 AU - Caitlin E. Mills AU - Kartik Subramanian AU - Marc Hafner AU - Mario Niepel AU - Luca Gerosa AU - Mirra Chung AU - Chiara Victor AU - Ben Gaudio AU - Clarence Yapp AU - Peter K. Sorger Y1 - 2021/01/01 UR - http://biorxiv.org/content/early/2021/08/28/2021.08.27.457854.abstract N2 - High throughput measurement of cell perturbation, by libraries of small molecules or gene knockouts, is a key step in functional genomics and pre-clinical drug development. However, it is difficult to perform viable, single-cell assays in 384-well plates, limiting many studies to simple well-average measurements (e.g. CellTiter-Glo®). Here we describe a public domain “Dye Drop” method in which sequential density displacement is used to perform multi-step assays for cell viability and EdU incorporation followed by immunofluorescence imaging. The method is rapid, reproducible, can be readily customized, and is compatible with either manual or automated laboratory equipment. We demonstrate Dye Drop in the collection of dose-response data for 67 drugs in 58 breast cancer cell lines and separate cytostatic and cytotoxic responses, thereby providing new insight into the effects of specific drugs on cell cycle progression and cell viability. Dye Drop substantially improves the tradeoff between data content and cost, enabling collection of large information-rich datasets.Competing Interest StatementThe authors have declared no competing interest. ER -