Problems with histotechnology

From:"Jasper, Thomas"

Hey Melanie,

I am a bit confused about the problem you are having with "water" under the
tissue which is showing up in staining.  If I understand you correctly you
believe that water is trapped under your tissue section, makes it all the
way through your staining procedure and shows up stained at the end.  If
this is the case it brings to mind the following questions:

1)	Are you H and E staining this tissue?
2)	Once your sections are cut do you place them in an oven or on a hot
plate to dry?
3)	If the answer is yes to #2 on your stain run down do you begin with
xylene (typically 3 stations)?
4)	Do you also have alcohol on the run down?  What percentage?
5)	How often do you change out your reagents?
6)	Is it possible that some of your stations are mixed up?  (i.e. A
xylene for an alcohol or vice-versa, or an alcohol which is not anhydrous
where one should be.)
Some of these questions may seem simple or obvious, but you may be barking
up the wrong tree.  I understand your concern with the charge on the slides,
but it sounds like you have eliminated that as a possibility.  I really have
a hard time believing that water could make it all the way through any
staining procedure when it is not supposed to be there.  Particularly if you
are drying and dehydrating your slides sufficiently, rehydrating, staining
and sufficiently dehydrating again.  Perhaps my suggestions will help or
maybe I don't understand and am all wet!  No pun intended.  I hope this
helps.  Good luck!

Thomas Jasper HT(ASCP)BAS
Anatomic Pathology Coordinator
SMDC Clinical Laboratory
Duluth, MN

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