From:"Dr. Ian Montgomery." <ian.montgomery@bio.gla.ac.uk>

<html> Su,<br> <x-tab>        </x-tab>Let me offer some advice before bowing to John's greater knowledge. <br> <x-tab>        </x-tab>Using acetone at -80C you are freeze substituting the tissue. Depending on size of tissue sample this can take many days - remember, small is beautiful. If possible use a rotator in the freezer or frequently agitate the tissue. Warm up slowly and evenly, probably over a day. <b>Do not use any solvents with water, your tissue is dehydrated so why hydrate again.</b> Methyl benzoate, mmmmm, wonder why, it is a clearing agent, not my first choice, but can be used to make tissue less brittle. Xylene is used here as a linking agent between methyl benzoate and wax. <br> <x-tab>        </x-tab>Depending on the tissue and size I'd be tempted, in the first instance, to have a trial. Wring the changes and see what happens, but definitely, <b>no water.<br> </b>Ian. <br> <br> <br> <blockquote type=cite class=cite cite>Date: Fri, 30 Mar 2001 07:51:22 -0500<br> From: "Su, Phy-Huynh" <psu@shctampa.usf.edu><br> To: "'J. A. Kiernan'" <jkiernan@uwo.ca>,<br>    "'Histonet'" <Histonet@pathology.swmed.edu><br> <br> Dear Histonetters:<br> <br> May I ask a question about methyl benzoate, please.<br> <br> I have received a protocol to fix tissues in acetone, at -80 degree C with<br> few changes, then bring the tissues up to RT in a fresh one, then going into<br> methyl benzoate before going into xylene, then paraffin.  I understand that<br> methyl benzoate is a clearing agent, as xylene, but is it really necessary?<br> Does it act as another clearing agent here or does it have any other<br> affects?  Can I just go directly to Xylene as routine processing?  How about<br> putting the tissues into 50%, 70%, 100% alcohol?  Will this deterioate the<br> tissues?<br> <br> Thank you.<br> Su</blockquote> <x-sigsep><p></x-sigsep> <font color="#0000FF">Dr. Ian Montgomery,<br> West Medical Building,<br> University of Glasgow,<br> Glasgow,<br> G12 8QQ.<br> Tel: 0141 339 8855.  Extn:6602.<br> Fax: 0141 330 2923<br> e-mail: ian.montgomery@bio.gla.ac.uk</font></html>
<< Previous Message | Next Message >>