[Histonet] automated IHC on bone sections?

From:"Garlits, John"


I am writing to ask if anyone out there has successfully done automated
IHC on decalcified, paraffin-embedded sections.  I have been staining
these for GFP, manually, for some time, but I have found that my IHC
results are quite variable.  I would like to work out an automated
method that might be faster and more consistent.  I have submitted
sections for automated staining at an in-house facility, but the problem
is that the actual bone part of my mouse femur sections always peels
off.  I use an enzyme-based antigen retrieval method when I do them
manually, but I understand that automated stainers typically use a
heat-based method.  I suspect that the heat-based retrieval is what
makes the bone peel off because I have tried a manual version of this
and witnessed the same result.  Do these automated stainers have the
ability to omit the heated retrieval step, or even pause it to introduce
an enzyme solution?

John Garlits
St. Jude Children's Research Hospital
Memphis, TN

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