Hi Louise

(... evil grin, to begin with ...). When I criticized the Dako waterbath
protocol as inadequate, I didn't get too much praise from Dako. However,
having our lab at 560 m above sea level (and not on a Californian sea
shore), we get our water boiling a little earlier than Dako Carpinteria. We
tried anything under the sun to get the temperature inside all sorts of
Coplin jars (glass, plastic, TPX) in a waterbath to a constant of 95 C
(minimum) ... and failed. Today we put the slides in a metal rack (vertical)
and put a constant number of 20 slides (blanks, as necessary) in a good old
Duran glass beaker and put it on a Bunsen burner, modulate the heat of the
flame - and get perfect results. The Dako rep visited our lab and was
impressed. Now, I have an advantage over most Histonetters: I live in
Europe, where Bunsen burners are not yet considered to be a modified sort of
atomic bomb, so we may still use them in our labs ... If open flames are
still tolerated in your lab, you may want to try our protocol (may be not
just next to the hood with the organic solvents ...). If open flames are a
no-mo, may be you can find a different source of heat that allows you to
bring the buffer in the beaker to boil, and which can rapidly be reduced, so
your buffer doesn't evaporate over the 40 min your slides are sitting in the
beaker. Good luck!

Andi Kappeler
Institute of Pathology, University of Bern, Switzerland