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Related Experiment Video

Updated: Sep 9, 2025

Spatial Profiling of Protein and RNA Expression in Tissue: An Approach to Fine-Tune Virtual Microdissection

Spatial Profiling of Protein and RNA Expression in Tissue: An Approach to Fine-Tune Virtual Microdissection

Published on: July 6, 2022

Alternate dyes for image-based profiling assays.

Suganya Sivagurunathan¹, Patrick Byrne², Alán F Muñoz¹

¹Imaging Platform, Broad Institute of MIT and Harvard, Cambridge, MA, USA.

SLAS Discovery : Advancing Life Sciences R & D

|August 30, 2025

Summary

This summary is machine-generated.

New dyes minimally impact Cell Painting assay performance, enabling enhanced cellular profiling. Live cell imaging with ChromaLive dye tracks morphological changes in real-time for expanded feature space.

Keywords:

Cell painting ChromaLive High content screening Image-based profiles Live cell painting MitoBrilliant Morphological profiling Phenotypic screening Phenovue phalloidin 400LS

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Optimized Staining and Proliferation Modeling Methods for Cell Division Monitoring using Cell Tracking Dyes

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Published on: December 13, 2012

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Last Updated: Sep 9, 2025

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Published on: July 6, 2022

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Optimized Staining and Proliferation Modeling Methods for Cell Division Monitoring using Cell Tracking Dyes

Optimized Staining and Proliferation Modeling Methods for Cell Division Monitoring using Cell Tracking Dyes

Published on: December 13, 2012

Area of Science:

Cellular imaging and analysis
High-content screening
Image-based profiling

Background:

Cell Painting is a key assay for image-based profiling, using six dyes to stain cellular compartments.
Profiles reveal connections between samples and assess treatment impacts.
Researchers may modify dyes for specific phenotypes or live-cell imaging.

Purpose of the Study:

Evaluate novel dyes for Cell Painting assay.
Assess dyes that replace or augment traditional stains.
Test live-cell compatible dyes for dynamic imaging.

Main Methods:

U2OS cells were treated with 90 compounds.
Cells were stained with standard Cell Painting dyes, or with MitoBrilliant or Phenovue phalloidin 400LS.
Live-cell ChromaLive dye was also tested.

Main Results:

All dye sets distinguished biological replicates from negative controls.
Distinguishing between different compound treatments was challenging for all sets.
Live cell dye showed distinct profiles across compound classes, with later time points being more informative.

Conclusions:

MitoBrilliant and Phenovue phalloidin 400LS substitutions had minimal impact on Cell Painting performance.
Phenovue phalloidin 400LS aids in isolating actin features and accommodates additional dyes.
Live cell imaging with ChromaLive dye allows real-time assessment of morphological changes, expanding profiling capabilities.