Dear Louise:

The Declere protocol calls for the following:

A) 3 micron thickness of sections on positively charged slides.
B) Dry 1 hour in an incubator which has been properly calibrated at 58 C.
C) Use either autoclave (metal pressure cooker with conventional heat
source), conventional steamer, or other conventional heat source.

Additional comments: If your sections are thicker (up to 4 microns maximum)
then dry longer - 2 hours at 58 C.  If you are dealing with breast tissue
which tends to be fatty, 3 or 4 hours drying may be necessary. We have had
good experience drying overnight at 38 C. Also worth mentioning is that not
all positively charged slides are alike - we have had great consistency
using Anapath Product No. 318. Finally, Declere works best on neutral
buffered formalin. Zinc formalin tends to produce sections that are more
difficult to cut and consequently more difficult to get to adhere to the
slides consistently. Many labs have had difficulty with this fixative even
in conjunction with conventional clearing and citrate unmasking.

If the above measures are carefully and consistently practiced, you will
virtually never have any tissue adhesion problems.

I will be happy to address any additional questions.

Regards,

Paul Ardi
Cell Marque Corp.
800-665-7284
512-306-8899



At 03:44 PM 11/23/98 +0000, you wrote:
>Hi histonetters,
>
>We are experiencing a problem with charged slides and "Declere"
>solution from Cellmarque. We find that a large proportion of tissue
>sections (formalin fixed, paraffin embedded) exposed to microwave
>pretreatment with this solution are washing off.
>("Declere" is purported to be a combination dewaxing, rehydrating and
>antigen retrieval solution)
>Has anyone else had this problem and is there a way to circumvent it?
>
>All comments will be welcome
>many thanks
>Louise Taylor
>SAIMR
>Johannesburg
>South Africa
>
>
>
>