It's Calcein AM / Ethidium homodimer-1 stains. I stained chondrocytes and the images I got show that the Calcein AM stained all the cells, even the ones stained with the Ethidium Homodimer-1.
I think the concentrations I used for the staining solution are not accurate.
Does these kit differentiate between viable and dead chondrocytes?
You should be able to use both stains at 1 uM final concentration in the cells. EthD1 is quite bright, and you may be able to get sufficient discrimination at 0.1 uM.
It's very important to note that calcein AM gets pumped out of cells by organic anion transporters. You should image your samples within 2-4 hours of staining.
Thank you very much Christina for the informative answer and suggestions. I will try as you said and firstly with the reduction in EthD and see how it goes from there.
Thanks again,
Alia
Thanks Jordan for the the information. I did image them within the 2 to 4 hour time frame, but I will adjust the concentrations of the EthD1 as suggested.
Thank you.
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