Dr. Weiler,
I use the live/dead kit in bovine and mouse cartilage studies and find that it
works nicely in a range of 200nM-2uM for calcein AM and 4-16uM for EthD-1.
I use tissue that has been in organ culture, so I wash thoroughly with PBS
before staining to remove any esterase activity present in the serum-containing
culture media.  These probes penetrate into the cartilage matrix well; in fact;
rather than using thin slices, I use sections of around 1mm3 and find that for
my purposes, the interior stains adequately. In thin sections you should have
no trouble at all.  Regarding incubation, I add the probes directly to the
culture medium (small volume) and incubate at 37 degrees for approximately half
an hour.

Kiley Ariail
Shriner's Hospital for Children
Portland, Oregon, USA

>Hello all,
>we want to study cell viablity in cartilage samples by the use of the
>live/dead Viability Kit (molecular probes calcein AM/ethidium
>homodimer). Does anybody have any experiences on this issue. We want to
>use thin slices of cartilage but according to the matrix we do not know
>anything about incubation or penetration of dyes or thickness of slices.

>Thank you very much in advance

>Dr. Christoph Weiler
>Office
>Hopital Cantonal Fribourg
>Departement Orthopedie
>CH-1708 Fribourg
>Switzerland
>Tel.         0041-26-426-7223
>Fax:         0041-26-426-7225
>E-Mail:        [log in to unmask]