Cell Viability Staining Kit

April 04, 2018

Nottingham Trent University
JvGCRC
Research Fellow

Overall

Quality of Results

Ease-of-Optimization

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Company:

Thermo Fisher Scientific

Product Name:

LIVE/DEAD™ Fixable Yellow Dead Cell Stain Kit

Catalog Number:

L34959

This stain has been used in a flow cytometry based study immunophenotyping PBMC populations obtained from cancer patients. The results have not yet been published. Multi-color antibody panels were designed to identify a number of lymphocyte populations and this stain was used to confirm cell viability.

Experimental Design and Results Summary

Application

Flow Cytometry

Starting Material

Human peripheral blood mononuclear cells (PBMC)

Protocol Overview

Kit is supplied as aliquots of stain that are prepared by reconstituting with the supplied DMSO. Stain was added to cells in PBS and left to incubate for 30 minutes at 4oC. Following titration and optimisation experiment it was found to be possible to reduce the suggested incubation volume to 100uL (from 1mL) and the stain could be added at the same time as the fluorescently-labelled antibodies hence saving time within the staining protocols. Cells can be analysed straight away or they can be fixed to allow further interrogation (intracellular staining for example).

Tips

Aliquot unused stain and store at -20oC for future use. Protect from light.

Results Summary

Very clear distinction of live and dead cells. Yellow stain is compatible with a little-used channel on the violet laser which helps when designing multi-colour flow cytometry panels.

DOI or PMID #

NA

Additional Notes

NA

Image Gallery

Summary

The Good

Simple protocol that provides very clear staining of non-viable cells. Supplying multiple aliquots of stain means little waste.

The Bad

Care is required when handling to ensure minimal exposure to light, and to minimise the number of freeze/thaw cycles.

The Bottom Line

Excellent product that is my 'go-to' viability stain for all flow cytometry protocols.

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