High-throughput method to make phenotypic analysis of human cells cultured in vitro
2016
Broad Institute of Harvard and MIT, Cambridge, USA(1)
Recursion Pharmaceuticals, Salt Lake City, USA(2)
Recursion Pharmaceuticals, Salt Lake City, USA(2)
High-throughput microscopy has been a particularly fruitful type of phenotypic screening. However, most large-scale imaging experiments extract only one or two features of cells, and/or aim to identify just a few ‘hits’ in a screen, meaning that vast quantities of quantitative data about cellular state remain untapped. In the present study, the researchers aimed at developing a method called Cell Painting assay, which is a generalizable and broadly applicable method for accessing the valuable biological information about the cellular state that is contained in morphology. The assay was developed with a range of widely available human cell lines and two primary cells all cultured in vitro. Cells are plated in multiwell plates, perturbed with the treatments to be tested, stained, fixed, and imaged on a high-throughput microscope. The researchers used six fluorescent stains revealing eight cellular components or compartments in a single microscopy-based assay. Automated image analysis pipelines extract ~1,500 morphological features from each stained and imaged cell to produce profiles. Profiles are then compared against each other and mined to address the biological question at hand. In conclusion, the study establishes a new method to make rich phenotyping at single-cell resolution in a high-throughput fashion.
Cell Painting, a high-content image-based assay for morphological profiling using multiplexed fluorescent dyes
Anne E Carpenter(1), Christopher C Gibson(2)
Added on: 12-22-2021
[1] https://www.nature.com/articles/nprot.2016.105
