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Bioactivity screening of environmental chemicals using imaging-based high-throughput phenotypic profiling

Metadata Updated: May 2, 2021

In the present study, we adapted an existing phenotypic profiling assay (“Cell Painting”, (Bray et al., 2016)) to be compatible with in-house microfluidics capabilities for 384-well culture format, chemical exposures and fluorescent cytochemistry in order to facilitate concentration-response screening of several hundred environmental chemicals. In this assay, human-derived cells were labeled with multiple fluorescent probes to visualize various subcellular organelles and structural features. High content image analysis workflows were used to measure hundreds of morphological features at the level of the individual cell (i.e. shape of the cells, intensity, texture and distribution of fluorescent labels, etc.). The resultant data were then used to calculate well-level summary values, perform high-throughput concentration-response modeling and generate phenotypic response profiles. First, we identified and screened a set of candidate phenotypic reference chemicals for use as plate-based controls for evaluating HTPP assay performance during large-scale screening studies and identified an optimal exposure duration for HTPP screening. Second, we screened a set of 462 environmental chemicals in the U-2 OS cell model and derived in vitro potency estimates for bioactivity of all active chemicals. In addition, we demonstrated the technical reproducibility of the HTPP assay in concentration-response screening mode using the previously identified phenotypic reference chemicals. Next, we used reverse dosimetry to calculate administered equivalent doses (AEDs) corresponding to the thresholds for chemical bioactivity and compared those values to in vivo effect values from mammalian toxicity studies.

This dataset is associated with the following publication: Nyffeler, J., C. Willis, R. Lougee, A. Richard, K. Friedman, and J. Harrill. Bioactivity screening of environmental chemicals using imaging-based high-throughput phenotypic profiling. TOXICOLOGY AND APPLIED PHARMACOLOGY. Academic Press Incorporated, Orlando, FL, USA, 389: 114876, (2020).

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Public: This dataset is intended for public access and use. License: See this page for license information.

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Metadata Created Date March 29, 2021
Metadata Updated Date May 2, 2021

Metadata Source

Harvested from EPA ScienceHub

Additional Metadata

Resource Type Dataset
Metadata Created Date March 29, 2021
Metadata Updated Date May 2, 2021
Publisher U.S. EPA Office of Research and Development (ORD)
Data Last Modified 2020-04-20
Public Access Level public
Bureau Code 020:00
Schema Version
Harvest Object Id 4a238fac-44e2-43fe-8e1d-9d7e472fece0
Harvest Source Id 04b59eaf-ae53-4066-93db-80f2ed0df446
Harvest Source Title EPA ScienceHub
Program Code 020:095
Publisher Hierarchy U.S. Government > U.S. Environmental Protection Agency > U.S. EPA Office of Research and Development (ORD)
Related Documents
Source Datajson Identifier True
Source Hash 05882e5e631a90c887f931040aad4f037ebfa9df
Source Schema Version 1.1

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