Dear Yi

Your loading time seems long. I would suggest cutting this back and
also not waiting more than 30 mins before imaging. The dye will be
lost from the cell as it is a living cell that will extrude/sequester
such a molecule. You could try something like probenecid but I've no
experience of this. We never have had to look at cells for very long
periods...


Regards

Mark Cannell

On Tue, 2 Sep 1997 18:09:30 +0100 Yi Cui wrote:

> From: Yi Cui <[log in to unmask]>
> Date: Tue, 2 Sep 1997 18:09:30 +0100
> Subject: loading cells with fluo3
> To: [log in to unmask]
>
> hi, all
>
> I am trying to record the Ca transient of guinea-pig heart cells
using
> fluo-3. I load the cells with fluo-3 AM 5-10uM for about 30-60min
and give
> 3 time washout. The signal is  quite good in the first one hour
after
> loading and then gradually become weak. It appears that the dye lost
from
> the cell. Anybody there have any experince or suggestion of keeping
the
> dye
> in the cell.
>
> thanks
>
> yi