RE: vascular perfusion

From:Cynthia Favara <cfavara@niaid.nih.gov>

<!DOCTYPE HTML PUBLIC "-//W3C//DTD HTML 4.0 Transitional//EN"> <HTML><HEAD> <META HTTP-EQUIV="Content-Type" CONTENT="text/html; charset=iso-8859-1"> <META content="MSHTML 5.50.4134.600" name=GENERATOR></HEAD> <BODY> <DIV><SPAN class=911281502-20032001><FONT face=Arial color=#0000ff size=2>Well yes I have perfused mice with Bouins. We perfuse white mice and they turn a lovely shade of yellow. I use the same protocol as perfusing with NBF. I flush with 5 ml of heparinized saline 30units/ml followed by fixative about 10ml depending on the size of the mouse. Trying to get 2-3 times the blood volume. I post fix overnight and them rinse in PBS until the rinse is clear. Usually takes multiple rinses. I do this in containers on a rotator. If I start in the morning the specimen is usually ready to process that evening. Let me know if I can be of further assistance.</FONT></SPAN></DIV> <DIV><SPAN class=911281502-20032001> <P><FONT face="Arial Narrow" color=#ff00ff size=2>Cynthia Favara</FONT> <BR><FONT face="Arial Narrow" color=#000080 size=2>Rocky Mountain Laboratories</FONT> <BR><FONT face="Arial Narrow" color=#000080 size=2>903 S. 4th Street</FONT> <BR><FONT face="Arial Narrow" color=#000080 size=2>Hamilton, MT 59840</FONT> <BR><FONT face="Arial Narrow" color=#000080 size=2>406-363-9317</FONT> <BR><FONT face="Arial Narrow" color=#000080 size=2>FAX 406-363-9286</FONT> </P></SPAN></DIV> <BLOCKQUOTE dir=ltr style="MARGIN-RIGHT: 0px"> <DIV class=OutlookMessageHeader><FONT face="Times New Roman" size=2>-----Original Message-----<BR><B>From:</B> Peter Poon [mailto:pcp512@hotmail.com]<BR><B>Sent:</B> Monday, March 19, 2001 11:17 AM<BR><B>To:</B> HistoNet@pathology.swmed.edu<BR><B>Subject:</B> vascular perfusion<BR><BR></FONT></DIV> <DIV> <P>Has anyone done vascular perfusion fixation with Bouins' for IHC in rodent<BR>brains?<BR><BR>If so<BR>1.)how much/long to prefuse?<BR>2.) post-prefusion fixation , if any? (time and temperature)<BR>3.) Wash - a) in what?<BR>            b) how long and what temp?<BR><BR>any advice is appreciated.<BR><BR>Peter<BR></P></DIV><BR clear=all> <HR> Get your FREE download of MSN Explorer at <A href="http://explorer.msn.com">http://explorer.msn.com</A><BR> <P></P></BLOCKQUOTE></BODY></HTML>
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