Louise:
  When I did IHC manually I used to rinse in the following way:
  In a glass container with FRESH (unused) PBS buffer, there was a slide holder; I removed the slides from the wet chamber after the given incubation period was completed and transferred the slides (one at a time) to the slides holders. Once all were there, I just moved the holder with the slides several times (5-7) and that was it.
  Took the slides out (one at a time), wipe them back the slides and around the tissue and added the following reagent.
  While incubating I discarded the used PBS and added fresh one for the next washing. Washing was OK without background and strong signal. If you want you could have 2 containers with PBS and after washing in the first could go to the second. Washing never took more than 1 minutes if 2 washing containers were used.
  Hope this will help you.
  René J.

louise renton  wrote:
  Hi all,

How long and/or vigorous should buffer rinses between immunohisto
steps be(ffpe 4um sections)? I am rinsing 5mins with agitation in PBS
+0.05% Tween. Is this too long? I am concerned that I am washing away
signal yet if I use PBS alone there is overwhelming background and
general muckiness

-- 
Louise Renton
Bone Research Unit
University of the Witwatersrand
Johannesburg
South Africa
"There are nights when the wolves are silent and only the moon howls".
George Carlin
No trees were killed in the sending of this message.
However, many electrons were terribly inconvenienced.

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