Great Fixable Viability Dye For Flow Cytometry

June 21, 2018

BlueskyReddit
Johns Hopkins School of Medicine
Oncology
Graduate Student

Overall

Quality of Results

Ease-of-Optimization

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Company:

Thermo Fisher Scientific

Product Name:

Fixable Viability Dye eFluor™ 780

Catalog Number:

65-0865-14

We are a T cell lab who conducts a lot of flow cytometry experiments. Since most of our analysis is done with tissue such as tumors or lungs, it is crucial we include a viablity dye to exclude non-viable cells from analysis. We use this fixable dye since preserving signal is important for our fixed samples. This is a very nice dye that fits into our flow panel using the Red laser of most cytometers. It is very easy to use, reliable, and somewhat inexpensive compared to other fixable dyes.

Experimental Design and Results Summary

Application

Flow cytometry

Starting Material

Single cells from murine tissue

Protocol Overview

We obtain single cell suspension of cells for flow staining. We first perform normal surface staining of antibodies. Next, we wash the cells with PBS to remove any protein, and then stain the cells with the viability dye for 20 minutes at 4 deg before fixation.

Tips

Titrate the dye before using since it can be quite bright depending on cytometer and lot of dye.

Results Summary

We are able to consistently discriminate live cells from dead cells during flow acquisition. The separation is quite nice to easily indicate viable cells.

DOI or PMID #

27064374

Additional Notes

N/A

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Summary

The Good

Great robust fixable viability dye

The Bad

None really. Just need to titrate before use to avoid bleeding of signal if too bright.

The Bottom Line

Great dye to exclude non-viable cells for flow cytometry. Fits nicely into our flow panels since we don't use any other markers in the APC-Cy7 channel. Inexpensive as well since titration sometimes is 1:5000, so one vial goes a long way.

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