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Multiplexed and reproducible high content screening of live and fixed cells using the Dye Drop method

Multiplexed and reproducible high content screening of live and fixed cells using the Dye Drop method

Authors :
Chiara Victor
Luca Gerosa
Ben Gaudio
Mario Niepel
Kartik Subramanian
Marc Hafner
Peter K. Sorger
Mirra Chung
Clarence Yapp
Caitlin E. Mills
Publication Year :
2021
Publisher :
Cold Spring Harbor Laboratory, 2021.

Abstract

High-throughput measurement of cells perturbed using libraries of small molecules, gene knockouts, or different microenvironmental factors is a key step in functional genomics and pre-clinical drug discovery. However, it remains difficult to perform accurate single-cell assays in 384-well plates, limiting many studies to well-average measurements (e.g. CellTiter-Glo®). Here we describe a public domain “Dye Drop” method that uses sequential density displacement and microscopy to perform multi-step assays on living cells. We use Dye Drop cell viability and DNA replication assays followed by immunofluorescence imaging to collect single-cell dose-response data for 67 investigational and clinical-grade small molecules in 58 breast cancer cell lines. By separating the cytostatic and cytotoxic effects of drugs computationally, we uncover unexpected relationships between the two. Dye Drop is rapid, reproducible, customizable, and compatible with manual or automated laboratory equipment. Dye Drop improves the tradeoff between data content and cost, enabling the collection of information-rich perturbagen-response datasets.

Details

Database :
OpenAIRE
Accession number :
edsair.doi...........e5c359da92cf23f9d735736ac8beec71
Full Text :
https://doi.org/10.1101/2021.08.27.457854