Hi Christine,

Before we automated we used to circle the section with petroleum jelly (
vaseline) to create the reagent well.

Run the slides down to water. Fill a 10cc syringe with jelly. Carefully wipe
around the section area to make it dry. Apply the jelly around the tissue with
the syringe. Take a wet tissue paper and press the outside edge of the jelly
against the slide to form the reagent well.This take a little practice. When
finished the staining, run the slides in hot water and the jelly floats off.
Counter stain with Heam. and coverslip as ususal.This is one of the "pearls of
wisdom" that I picked up from a Hamilton , Ontario , Canada researcher in a
hallway discussion at the NSH convention a few years back. They used very high
dilution of a their homemade antibody to conserve on the antibody and
incubated in the frig for 2 days. By placing a coverslip on the vaseline she
created a humidity chamber which allowed the section to remain moist during
the incubation.

Sincerely,
Dan Botsford
Windsor Regional Hospital
Windsor, Ontario, Canada

cmcbride@tulane.edu wrote:

> Hi,
> Does anyone have a better alternative to PAP pens? Their membrane seems to
> breakdown after a few soln changes.
> Thanks,
> Christine
>
> Christine McBride, M.S.
> Tulane University Medical Center
> Center for Gene Therapy
> SL-99
> 1430 Tulane Ave.
> New Orleans, LA 70112
> cmcbride@tulane.edu
> Phone:(504) 988-7069
> Fax:(504) 588-5326