3 replies to this topic

[labnewbie]

Is it possible to stain with trypan blue in the petri dish itself, without lifting the cells and counting them in a hemacytometer?  If so, does anyone have a protocol for it?

[scolix]

how do you plan to get the cells into hemocytometer without lifting them?

Just add a few microliter of trypan blue will stain your cells on a petri dish. But you have to trypsinize them to get them into a hemocytometer.

[labnewbie]

that's the thing, i don't want to get them into a hemocytometer--I don't want to lift or count in a hemocytometer.
I want to be able to stain them in the dish so I can just visualize approx number of dead cells, after doing that, I want to stain the cells.  I want to be able to distinguish between dead and alive cells.
Do you have any suggestions/protocol of dilution of trypan blue/how long to leave in the dish. have you ever done it before?

[scolix]

I havn't stained for trypan and with another staining simultaneously. You could try it. But wash well, before the actual staining procedure.

For the trypan staining, I just add 10ul into 100ul cell suspension and you can look at them immediately under the hemacytometer.

One concern would be that during the staining procedure, you could lose the dead cells.
May be setup 2 dishes, one for counting dead cells with trypan and other for staining.