[Histonet] drosophila sectioning

[Anthony Gatt]

Thanks, Rene.  Any advice on a softening agent and where it would fit into the protocol?

---- Original message ----
>Date: Fri, 11 May 2007 09:15:23 -0700 (PDT)
>From: Rene J Buesa <rjbuesa <@t> yahoo.com>  
>Subject: Re: [Histonet] (no subject)  
>To: Anthony.Gatt <@t> jefferson.edu, "histonet <@t> lists.utsouthwestern.edu" <histonet <@t> lists.utsouthwestern.edu>
>
>   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 <Anthony.Gatt <@t> jefferson.edu> 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,
>
>     Anthony
>
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