Re: [Histonet] (no subject)

From:Rene J Buesa

Your problem resides in the fact that you have to "soften" the chitin before processing the heads, otherwise the chitin cannot be infiltrated by the paraffin, causing the tears you are having.
  Soften the chitin first and process afterwards.
  René J.

Anthony Gatt  wrote:
  Hello, I am currently sectioning drosophila heads and am tearing up the paraffin. Without the heads, I get beautiful ribbons so I suspect it is the sample itself. I am using a tissue processor with the following protocol after an o/n fix in 4% PFA.

15m -alcoholic formalin (x2)
30m -95% EtOH (x2)
30m -100% EtOH (x2)
30m -Histoclear (x3)
30m -Paraffin (x2)
45m -Paraffin

Embed immediately in paraffin, cold block 30m, section next day.

I am new to this an would greatly appreciate any help.

Thank you,


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