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

Caitlin E. Mills, Kartik Subramanian, Marc Hafner, Mario Niepel, Luca Gerosa, Mirra Chung, Chiara Victor, Benjamin Gaudio, Clarence Yapp, Ajit J. Nirmal, Nicholas Clark and Peter K. Sorger ()
Additional contact information
Caitlin E. Mills: Harvard Medical School
Kartik Subramanian: Harvard Medical School
Marc Hafner: Harvard Medical School
Mario Niepel: Harvard Medical School
Luca Gerosa: Harvard Medical School
Mirra Chung: Harvard Medical School
Chiara Victor: Harvard Medical School
Benjamin Gaudio: Harvard Medical School
Clarence Yapp: Harvard Medical School
Ajit J. Nirmal: Harvard Medical School
Nicholas Clark: Harvard Medical School
Peter K. Sorger: Harvard Medical School

Nature Communications, 2022, vol. 13, issue 1, 1-18

Abstract: 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.

Date: 2022
References: View references in EconPapers View complete reference list from CitEc
Citations:

Downloads: (external link)
https://www.nature.com/articles/s41467-022-34536-7 Abstract (text/html)

Related works:
This item may be available elsewhere in EconPapers: Search for items with the same title.

Export reference: BibTeX RIS (EndNote, ProCite, RefMan) HTML/Text

Persistent link: https://EconPapers.repec.org/RePEc:nat:natcom:v:13:y:2022:i:1:d:10.1038_s41467-022-34536-7

Ordering information: This journal article can be ordered from
https://www.nature.com/ncomms/

DOI: 10.1038/s41467-022-34536-7

Access Statistics for this article

Nature Communications is currently edited by Nathalie Le Bot, Enda Bergin and Fiona Gillespie

More articles in Nature Communications from Nature
Bibliographic data for series maintained by Sonal Shukla () and Springer Nature Abstracting and Indexing ().