[Histonet] Troubleshoot on mounting tissue

From:"Judy Choi"


This is my first time using this listserve, so I hope this works.

I have a question about mounting tissues (PFA perfused rat brain tissues to 
be specific).  I processed them for IHC using ABC complex system and DAB and 
used deionized water as my mounting medium.  However, when I put the tissue 
in water, it swells tremendously and easily becomes wrinkled.  I think as a 
consequence of the swelling, there are holes throughout my tissues.  (I 
didn't use hydrogen peroxide until DAB, and it was at an extremely low 
concentration of less than 0.1%.)

Anyways, is my hypothesis plausible (that mounting my tissue in DI water can 
cause tissues to swell and create holes)?  If so, does anyone know how I can 
eliminate this problem?  Or can it be another factor (bad cryoprotectant, 
bad perfusion, bad buffer)?  I appreciate any feedback I can get.

Thank you!

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