Using Cell Painting for Morphological Changes

To understand the cytological profiles of cells, a cell painting assay can be used to identify the difference in morphology of cell organelles. As the world of therapeutic drugs and technology advances, the cell painting approach helps in capturing the drugs reactions at a cellular level. This technique is a high-content, multiplex, image-based assay.

Cell painting is utilized for cytological profiling meaning a drug can be assessed by “painting” treated and non-treated groups of cells with non-overlapping stains that target different cellular compartments. This allows for scientists to visualize changes in different areas simultaneously, as well as track multiple pathways in parallel. Various organelles are painted with specific stains to identify them differentially to visualize phenotypic, physiological, metabolic, or epigenetic changes within each cell allowing large amounts of data to be compiled on how particular therapeutics operate and affect the samples. Morphological changes are assessed through an automated image analysis software, where each cell’s measurements will be identified. Measurements typically include intensity, texture, shape, and size as well as the proximity of an object to its neighboring structure which provides an indication of the spatial relationship between organelles.

Representative Composite image consisting of Red: Actin and golgi; Green: ER; and Blue: nuclei

Representative Endoplasmic reticulum
Dye: Concanavalin A/Alexa Fluor 488 conjugate

The cell painting assay can help the drug discovery world discern whether therapeutics are effective or not, and this assay can help to achieve this knowledge faster than in vivo or clinical studies. In addition, it can be used to characterize healthy cells from diseased cells. The scientists at Visikol use these techniques to identify the workings of many therapeutic agents, commonly with cancer cells. Recently, Visikol has been using cell painting to observe the epigenetic effects of therapeutics on cancer cells. In short, epigenetics refers to the effects on gene expression, which can be crucial to finding the right therapeutic for a disease. The cell painting assay has been utilized at Visikol for epigenetics experiments to measure the DNA methylation patterns using multiple methylation inhibitors to observe the effects on the cell characteristics.

Future plans include the assessment of two inhibitors of DNA methylation, 5-Azacytidine and 5-Deoxycytidine. 5-Azacytidine was first discovered almost 40 years ago and has shown various metabolic changes to cancer cells. Because it also has cytotoxic effects, this led to widespread use of 5-Deoxycytidine to elucidate the correlation between loss of methylation in specific gene regions and genes associated activation. Due to the widespread use, there has been an increased use of Decitabine as a therapeutic agent for cancers in which epigenetic silencing of critical regulatory genes has occurred. To avoid situations like this, it is important to gain a better understanding of how therapeutics work. Cell painting can give the maximum amount of information on multiple pathways and visual changes all through one assay.

If you are interested in learning more about this assay or any of the other many new research opportunities at Visikol, please reach out to our team. We are always interested in working together with our clients as a team to develop customized assays to best suit their needs.

Representative Nucleoli, cytoplasmic RNA
Dye: SYT0 14 green fluorescent nucleic acid stain

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