Great Dye for Tracking T Cell Proliferation

October 12, 2021

BlueskyReddit
Emory University
Pediatrics
Graduate Student

Overall

Quality of Results

Ease-of-Optimization

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Company:

ThermoFisher

Product Name:

CellTrace Violet

Catalog Number:

C34557

This dye was used to analyze T cell proliferation in different conditions. Cells are stained prior to culturing and lose half of the CellTrace Violet with each division, resulting in discrete peaks to show how many times the cells divided in 72 hours.

Experimental Design and Results Summary

Application

Flow cytometry

Starting Material

Blood T cells

Tips

if using PBMCs instead of purified lymphocytes, be sure to gate out unwanted cell populations to view clean peaks

Results Summary

This dye was used to analyze T cell proliferation in different conditions. Cells are stained prior to culturing and lose half of the CellTrace Violet with each division, resulting in discrete peaks to show how many times the cells divided in 72 hours.Results: The dye works well for identifying rounds of proliferation in stimulated lymphocytes. CD8+T cells that received anti-CD3 and anti-CD28 stimulation proliferated ~4 times in 72 hours, compared to freshly purified and cultured unstimulated cells which showed one uniform peak indicating no proliferation occurred.

DOI or PMID #

N/A

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Summary

The Good

clean results if you have optimized culturing conditions

The Bad

staining with this product does add in an extra step before you can actually start the assay, rather than doing all the staining at the end

The Bottom Line

great product if you have a good understanding of the conditions needed to achieve reliable proliferation

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