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Fixation and Sectioning

Fixation and Sectioning

Two basic types of preparation are used to visualize specimens with a light microscope: wet mounts and fixed specimens.
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Related Experiment Video

Updated: Jun 17, 2026

Cellular Redox Profiling Using High-content Microscopy

Cellular Redox Profiling Using High-content Microscopy

Published on: May 14, 2017

Live-cell painting: Image-based profiling in live cells using acridine orange.

Fernanda Garcia-Fossa^1,2, Thaís Moraes-Lacerda¹, Mariana Rodrigues-da-Silva¹

¹Departamento de Bioquímica e Biologia Tecidual, Instituto de Biologia CP 6109, Universidade Estadual de Campinas (UNICAMP) 13083-970, Campinas, SP, Brasil.

Molecular Biology of the Cell

|May 6, 2025

Summary

Live Cell Painting (LCP) offers a new way to study cell health in real-time without fixation. This high-content imaging method uses Acridine orange to track cellular changes, aiding drug and nanoparticle assessment.

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Last Updated: Jun 17, 2026

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Area of Science:

Cell biology
High-content imaging
Drug discovery

Background:

Image-based profiling is crucial for analyzing cellular responses to various treatments.
Current methods like Cell Painting require cell fixation, limiting live-cell analysis.
There is a need for live-cell imaging assays to assess cellular health and treatment effects dynamically.

Purpose of the Study:

To introduce Live Cell Painting (LCP), a novel high-content imaging method for real-time cell analysis.
To demonstrate LCP's utility in monitoring cellular responses to inhibitors, nanoparticles, and drug treatments.
To establish LCP as an accessible and versatile tool for cell health screening and biological investigation.

Main Methods:

Developed Live Cell Painting (LCP), a high-content assay utilizing Acridine orange, a metachromatic dye.
Applied LCP to track acidic vesicle redistribution in response to specific inhibitors.
Utilized LCP to detect subtle cellular changes induced by silver nanoparticles, outperforming MTT assays.
Employed LCP for dose-response assessments and clustering of drugs causing liver injury.

Main Results:

LCP successfully monitored dynamic changes in acidic vesicle redistribution.
LCP identified cellular alterations caused by silver nanoparticles, which were missed by MTT assays.
LCP enabled accurate dose-response evaluations and drug clustering based on cellular profiles, including liver injury.
The method proved effective in assessing overall cell health and treatment effects.

Conclusions:

Live Cell Painting (LCP) is an affordable, easy-to-use, image-based assay for assessing cell health.
LCP provides dynamic insights into cellular responses to drugs and nanoparticles.
LCP shows significant promise for initial cell health screening and addressing novel biological questions.