[Histonet] IHC Slides, Hotplate vs Oven

[Gagnon, Eric]

Tony's answer (62 degrees C for 25 mins) is the minimum to ensure section adheres to slide.  We all have to stain slides that have been baked longer than 30 mins.  However, especially for Her2, longer baking times can result in false negatives or at least diminished staining.  The justification for baking longer is an attempt to prevent the section washing off or lifting.  We are seeing evidence that section adhesion is not a good tradeoff for diminished staining.  Ideally both can be achieved with no tradeoff.
 
Tony or others who may wish to answer:
 
1. How do you monitor the shortened baking time, especially when multiple racks of slides are cut throughout the day?
2. Assuming charged slides and cell conditioning/antigen retrieval are employed, are you seeing major problems with section adhesion?
3. What if slides are received from another institution for staining, or slides get baked longer than 25 mins?
 
Thanks in advance,
 
Eric Gagnon MLT
Histology Laboratory
Kingston General Hospital
Kingston, Ontario, Canada