BioLegend
Zombie NIR™ Fixable Viability Kit
423106
RUO
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The Zombie NIR viability kit works well for identifying dead cells from cell suspensions isolated from mouse lungs, spleen, and lymph nodes. I recommend storing at -20C to ensure the dye works well over multiple freeze-thaw cycles. Do not store at 4C. Despite being excited by the red laser, this dye has worked for me even with APC, APC-fire750, and APC-810 in the same panel.
Flow cytometry
Mouse lung, mediastinal lymphnode, spleens
1:1000, 20 minutes at 4 degrees C in the dark
Fc Shield
BD cytofix/cytoperm; 100 ul for 20 mins at 4 degrees C
None
5 laser Aurora Flow cytometer
I used this product to identify Live (Zombie NIR negative population) immune cells from cells isolated from the lung, spleen and lymph nodes of mice. Cells were isolated from BALF using 1 ml of 1% BSA in PBS and from lungs through collagenase digestion; RBC lysis was performed using ACK buffer for all samples. Cells were then stained with viability dye in Fc block followed by extracellular antibodies such as CD45, TCR beta, CD4, CD8, and so on. After fixing and permeabilizing, I have also stained for intracellular cytokine expression. I have been impressed by the ability of the aurora to distinguish this dye’s signal (at R6) from that of APC-fire 750 (R7) and APC-fire 810 (R8).
N/A
Cheap and good signal.
Must be stored at -20 degrees C; takes time to thaw.
I would highly recommend this live-dead stain since it uses a channel that is highly compatible with most other common fluorophores.