Time resolved cell painting enables rapid assessment of cell phenotypes

Cell Painting is an advanced imaging technique for drug discovery used to study cellular phenotypes by simultaneously labeling various organelles/structures and analyzing the resulting multidimensional phenotypic features through a sophisticated data analysis pipeline. Based on established phenotypi...

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Main Authors: Franziska A. Hecker, Bruno Leggio, Tim Koenig, Karsten Niehaus, Sven Geibel
Format: Article
Language:English
Published: Elsevier 2025-09-01
Series:SLAS Discovery
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Online Access:http://www.sciencedirect.com/science/article/pii/S2472555225000425
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author Franziska A. Hecker
Bruno Leggio
Tim Koenig
Karsten Niehaus
Sven Geibel
author_facet Franziska A. Hecker
Bruno Leggio
Tim Koenig
Karsten Niehaus
Sven Geibel
author_sort Franziska A. Hecker
collection DOAJ
description Cell Painting is an advanced imaging technique for drug discovery used to study cellular phenotypes by simultaneously labeling various organelles/structures and analyzing the resulting multidimensional phenotypic features through a sophisticated data analysis pipeline. Based on established phenotyping methodologies, this method has relied on incubation times of typically around 48 h for the assessment of phenotypic fingerprints. Here we provide evidence that earlier assessments show more robust results with increased significance of phenotypic fingerprints that better reflect primary physiological effects.Our study included compounds that range from representatives with modes of action that result in immediate phenotypic changes, such as energy metabolism inhibitors, to representatives that typically show pronounced phenotypes after several days, such as developmental inhibitors. Remarkably, we observed that for all compounds, primary cellular alterations were best detected at early timepoints after treatment, specifically at 6 h for Sf9 insect cells and shortly later timepoints for mammalian U2OS cells. Brief incubation periods enable the capture of primary effects of treatments while minimizing the influence of secondary changes as well as downstream phenotypic alterations like, for example, cell death. This enhances the specificity and accuracy of Cell Painting and consequently provides a more immediate depiction of primary actions from compounds. Notably, it also improves the efficiency of experimental workflows.In conclusion, we propose a more rapid assessment of cell phenotypes and morphology in the Cell Painting assay to enable a higher throughput in drug discovery screenings.
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spelling doaj-art-fa348231928c4b6c922fd688734f0d522025-08-20T03:50:49ZengElsevierSLAS Discovery2472-55522025-09-013510024910.1016/j.slasd.2025.100249Time resolved cell painting enables rapid assessment of cell phenotypesFranziska A. Hecker0Bruno Leggio1Tim Koenig2Karsten Niehaus3Sven Geibel4University Bielefeld, Proteome and Metabolome Research, Bielefeld, GermanyR&D Disease Control, Bayer SAS, Crop Science Division, Lyon, FranceR&D Image-based Screening Systems, Bayer AG, Pharma Division, Wuppertal, GermanyUniversity Bielefeld, Proteome and Metabolome Research, Bielefeld, GermanyR&D Hit Discovery, Bayer AG, Crop Science Division, Monheim, Germany; Corresponding author.Cell Painting is an advanced imaging technique for drug discovery used to study cellular phenotypes by simultaneously labeling various organelles/structures and analyzing the resulting multidimensional phenotypic features through a sophisticated data analysis pipeline. Based on established phenotyping methodologies, this method has relied on incubation times of typically around 48 h for the assessment of phenotypic fingerprints. Here we provide evidence that earlier assessments show more robust results with increased significance of phenotypic fingerprints that better reflect primary physiological effects.Our study included compounds that range from representatives with modes of action that result in immediate phenotypic changes, such as energy metabolism inhibitors, to representatives that typically show pronounced phenotypes after several days, such as developmental inhibitors. Remarkably, we observed that for all compounds, primary cellular alterations were best detected at early timepoints after treatment, specifically at 6 h for Sf9 insect cells and shortly later timepoints for mammalian U2OS cells. Brief incubation periods enable the capture of primary effects of treatments while minimizing the influence of secondary changes as well as downstream phenotypic alterations like, for example, cell death. This enhances the specificity and accuracy of Cell Painting and consequently provides a more immediate depiction of primary actions from compounds. Notably, it also improves the efficiency of experimental workflows.In conclusion, we propose a more rapid assessment of cell phenotypes and morphology in the Cell Painting assay to enable a higher throughput in drug discovery screenings.http://www.sciencedirect.com/science/article/pii/S2472555225000425Phenotype progressionCell paintingMicroscopySpodoptera frugiperda Sf9U2OS Cells
spellingShingle Franziska A. Hecker
Bruno Leggio
Tim Koenig
Karsten Niehaus
Sven Geibel
Time resolved cell painting enables rapid assessment of cell phenotypes
SLAS Discovery
Phenotype progression
Cell painting
Microscopy
Spodoptera frugiperda Sf9
U2OS Cells
title Time resolved cell painting enables rapid assessment of cell phenotypes
title_full Time resolved cell painting enables rapid assessment of cell phenotypes
title_fullStr Time resolved cell painting enables rapid assessment of cell phenotypes
title_full_unstemmed Time resolved cell painting enables rapid assessment of cell phenotypes
title_short Time resolved cell painting enables rapid assessment of cell phenotypes
title_sort time resolved cell painting enables rapid assessment of cell phenotypes
topic Phenotype progression
Cell painting
Microscopy
Spodoptera frugiperda Sf9
U2OS Cells
url http://www.sciencedirect.com/science/article/pii/S2472555225000425
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AT karstenniehaus timeresolvedcellpaintingenablesrapidassessmentofcellphenotypes
AT svengeibel timeresolvedcellpaintingenablesrapidassessmentofcellphenotypes