I'm hoping to stain two cell populations then mix them together. I'm aware that crystals will mosaically label cell populations, I'm hoping to stain all cells to identify which population they are from.
These are HaCaT cells, should I trypsinize them, then add a high concentration to the suspension, spin them down, wash, and then replate together?
What would be a high concentration?
You can use Fuse-It-color from ibidi. Its a membrane fusion technology to incorporate molecules very efficient into cells.
Nils, I'll look into that, thank you, but we already have a ton of DiI so I think my PI is going to want me to use what we have...
Hi Phil,
is there any reason why you have to use DiI? As far as I know it's lipophilic and it will may go from cell to cell (if i remember correctly from some experiments I did in chicken).
Why not transfecting with a GFP and an RFP plasmid and mix them together? At least in zebrafish, people do this all the time.
Best of lucks!
J.
Hey man!
It was in cell culture, creating a stable cell line of GFP and RFP cells in my lines would take a long time. Transient transfections wouldn't label all the cells.
The dye didn't mix much at all. It is lipophylic, so removing the ethanol solution and crystals left only the label in the cells themselves, they didn't bleed much. I was worried about apotptoic fragments getting taken up, but it was a short mixing period, only overnight.
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