Great Fixable Viability Dye

May 09, 2017

Dana-Farber Cancer Institute
Cancer Immunology and Virology
Research Fellow

Overall

Quality of Results

Ease-of-Optimization

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Company:

Tonbo Biosciences

Product Name:

Ghost Violet 510

Catalog Number:

13-0870-T100

Human T cells were cultured for 4 days with monocyte-derived dendritic cells (moDCs) treated with LPS. After 4 days, T cells were harvested, stained with Ghost Violet 510 and fixed with 2% paraformaldehyde. Subsequently viability-stained and fixed cells were stained for various cell surface and intracellular markers. Ghost Violet 510 was used as it gives a good separation compared to other fixable viability dyes and also because it is quite economical.

Experimental Design and Results Summary

Application

Flow Cytometry

Starting Material

Human T cells

Protocol Overview

T cells were stained with a 1:800 dilution of Ghost Violet 510 prepared in sterile 1X PBS for 20 mins at RT. After this incubation, cells were washed with PBS containing 2% FBS and 2mM EDTA. Cells were then fixed with 2% PFA for 15 mins at 4 C. Fixed cells were then stained for intracellular markers.

Tips

Staining at RT seems to work better than at 4 C for this product.

Results Summary

Good separation of viable and dead cells was observed.

DOI or PMID #

N/A

Additional Notes

N/A

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Summary

The Good

Economical, can be used at a relatively high dilution, and easy to optimize for a particular cell type.

The Bad

Spills somewhat in to the Pacific Blue channel but can be compensated with proper controls.

The Bottom Line

Good option for the 510 channel on the violet laser and far more economical for similar kinds of reagents from other vendors.

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