First of all, 2-3 minutes in PBS to wash out the Michell solution is too littler. I used to wash it on an agitation platform for 30 minutes. After that another wash and into distilled water and, believe it or not, I did not use OCT.
  In the cold chuck on the cold bar of the cryostat, I placed a small drop of distilled water, placed the biopsy (cut in half if it was a punch Bx, or on edge it is was a shave Bx), INTO de drop of water and very gently froze the drop of water, along with the Bx, with bursts of the Histofreeze. It worked wonders, and I did not have to remove the OCT afterwards.
  After many years doing it this way, I started using OCT and in the same way; OCT on the frozen chuck, Bx inside the OCT and freezing both with the coolant bursts.
  BUT, for you, at this moment, first try increasing the Michell washing time.
  René J.

eileen dusek  wrote:
  Hi Everyone,
I have been in Histology for a looong time and have met my match. I recently have new duties involving kidney bxs. We receive a formalin, EM, and Michels (Zuess) fixed specimens.
The Formalin and EM bx are not an issue, the Michels is kicking by butt!
I transfer the specimen from Michels to PBS, to rinse the fixative, for about 2-3 mins of agitation. After the PBS the specimen goes into the first OCT. This is swirled for another 2-3mins. I repeat this process again.
My problem is I get specimens that have too much freezing artifact. The structures are "blown up" and not compact.
I appreciate any suggestions to help the problem Thanks

Eileen Dusek


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