Hello Paul,

From your message I understand that you plan to do time series
experiments with FITC labelled cells. I tried to study apoptosis with
FITC labelled anexin and found out that after the first few images
the cells were dying by necrosis. I found that exiting the
fluorescent dye a few times is enough to kill the cells. You probably
are very limited in the number of images you make.

Greetings,
Gert van Cappellen

> I was wondering if anyone could suggest a method of counting cells in a
> confocal field of view for use in an FITC-labelled-drug uptake study in
> keratinocytes. I'm finding that I need a method of counting the total number
> of cells, since some cells take up the drug very well and others don't at
> all, so its hard to estimate the proportion of cells fluorescing etc.
>
> I'm now using a two channel system so dual labelling is a possibility (eg
> with Texas Red conjugated to a live cell marker?), but I was hoping that
> there was a simpler solution, particularly since these are time course
> experiments over up to 24 hours.
>
> Thanks in advance!
>
> Paul
>
> Dr Paul White
> Research Officer
> Dermal Therapeutics R&D Programme
> Royal Children's Hospital
> Melbourne, Australia
>
>
> Loretta Donders
> Project Management Assistant
> Dermal Therapeutics R&D Programme
> The Royal Childrens Hospital
> Flemington Road
> Parkville   Vic   3052
> AUSTRALIA
> Phone:  61 3 9345 7909
> Fax: 61 3 9347 7763
> E-Mail:  [log in to unmask]
>

 ========================================================================
Gert van Cappellen, [log in to unmask] Erasmus University,
Endocr.& Reprod., Dr. Molewaterplein 50, 3015 GE Rotterdam, The Netherlands
Fax: +31 10 4366832