[Histonet] brain sections falling off slides during IHC

[Thurby, Christina]

Hello all,
Can any of you all with experience immunostaining brain sections please help.  My sections are almost completely washing off the slides.  I am using 10% NBF fixed canine brain sections.  Embedded in paraffin and sectioned at 4 microns on plus slides with distilled water in the water bath.  The slides are dried at 65 C for 15 minutes and allowed to stand overnight.  I am using a Dako Autostainer for staining.  There is no antigen retrieval step.  My staining procedure is approximately 3.5 hours long.  At the completion of the procedure, the tissue on my slides is 50-80% washed off.

Any thoughts??
Christina Thurby