LIVE/DEAD™ Near-IR Stain Enables Viable Cell Sorting by FACS

June 09, 2026

BlueskyReddit
State University of New York at Buffalo
Physiology and Biophysics
Postdoctoral Researcher

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Quality of Results

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Company:

Thermo Fisher Scientific / Invitrogen

Product Name:

LIVE/DEAD™ Fixable Near-IR Dead Cell Stain Kit, for 633 or 635 nm excitation

Catalog Number:

L10119

We use the LIVE/DEAD™ Fixable Near-IR Dead Cell Stain Kit, for 633 or 635 nm excitation, prior to FACS sorting to enable sorting of viable astrocytes. This allows us to exclude non-viable cells during sorting, which improves both the quality and interpretability of our downstream data, such as RNA-seq.

Experimental Design and Results Summary

Application

Fluorescence-Activated Cell Sorting

Starting Material

Dissociated brain tissue (e.g., hippocampal tissue) after myelin immunomagnetic removal

Tips

We incubate the cells with reconstituted reactive dye for 30 min at 4 °C, protected from light, and wash with FACS staining buffer before FACS sorting.

Results Summary

We use LIVE/DEAD™ to isolate viable mouse brain astrocytes during FACS sorting for subsequent RNA-seq analysis. We validated the kit by intentionally killing cells through boiling. The dye reacted with free amines both inside the cells and on the cell surface, resulting in intense fluorescent staining (shown in pink in the image).

DOI or PMID #

N/A

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Summary

The Good

Works consistently well and allows sorting of only viable cells.

The Bad

To prepare the dye, we need to add 50 µL of DMSO to the reactive dye in the vial. Usually, we use up to 10 µL, and the remaining amount is wasted (it can be used only within a few hours of reconstitution and cannot be stored).

The Bottom Line

Easy to use and works consistently well. We recommend this kit.

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