[Histonet] Section adherence issues (IHC)

[Gagnon, Eric]

Hi Greg,

Permit me a few thoughts on factors not mentioned in your email:

  *   Do your slides 'look' baked/is the wax melted when they are removed from the oven?
  *   Also, is there variance in your oven temperature?
  *   Especially vexing is the fact that control sections are lifting. Are the control sections pre-baked or at time of cutting the ER/PR patient sections?
  *   Are the sections completely dry before covertiles are placed on top of the slides in the Leica staining rack?
  *   Has Leica checked the instrument to rule out any instrument-related issues? Is the poor adherence occurring in one particular location within the instrument, or in various slide positions?

Fortunately, special stains and routine staining rarely seem to be affected by the various pH/mechanical/detection system variables that exist in IHC.

In our experience with Bond-III, areas of poor adherence tend to be longitudinally lifting/up 'the middle' of the slide and are usually resolved by leaving slides of only such problem tissues (as you mentioned) for longer times in the oven than normal.

Hope this helps,
Eric Gagnon MLT
Kingston General Hospital
Kingston, ON



We have a Bond-III immunostainer and we use the Leica Apex charged slides
for our IHC stains. We have no additives in our water baths. Our sections
drain in a stand and then any trapped water under the section is either
flicked or wicked away as needed prior to placing the slides in a rack in a
60 C oven for 30 mins prior to staining.  We do not do on board baking
(primarily because on board baking is only 10 mins long and with the
section lifting issue we want longer).

Some of the specimen types are more susceptible it would seem. Cervix LEEP
specimens tend to be bad, we use a 4mm punch to obtain some of our control
tissue and so the breast tissue we use for Myosin Heavy Chain seems to be a
bad one and sometimes our ER/PR control sections (but not always). Fine
needle cores where a lot of tumour is present tend to be bad (again not
always).

We have tried baking longer, we turn ourselves inside out trying to get all
of the water out from under the sections, we have tried charged slides from
another manufacturer. I have looked at the HIER protocols and none are
extraordinary in nature. We use very clean covertiles (no scratches or
blemishes). Our specimens are fixed for 24hrs before processing. We use the
same slides for Special Stains and don't have this issue there.

I need some new suggestions to try! All ideas welcome.
Cheers,
Greg