[Histonet] Eosin Leaching

[fivetoads <@t> charter.net]

We are having an intermittant but ongoing problem with our routing hematoxylin and eosin (H&E) stains leaching out the eosin after coverslipping anywhere from 1 hour to 1 day later.  We use Bio-Care Cat-Hematoxylin and their eosin with phloxine, which is new to us this year.  I do not think this brand has anything to do with the problem, as we have experienced this before.  We use 4 absolute ethyl alcohol changes after the eosin for a total dehydration time of 6 minutes.  After that we have 3 changes of Slide Brite (xylene substitute that we have used for the past 8 years)for a total clearing time of 6 minutes. This is all done on an automated stainer.  We 'dump and rotate' the absolute alcohol solutions after every basket of 20-30 slides.  We dump and rotate the clearing agent 2 times per week, more often when the slide count increases.  (100 slides a day is a busy day for our small lab).  We use a mounting medium  that is designated for use with the clearing agent.  All of!
  our ducks seem to be in 'a row'...what am I missing here??

Jonell
Bay Area Hospital
Coos Bay, Or