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<TITLE>[Histonet] Mice embryonic brain - EM Poor tissue quality - need help</TITLE>
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<DIV>I'm not sure about EM procedures, but I've been told, and do find,
perfusion fixation to be the best for fixing brain tissue. If you need a
protocol for that I have one.</DIV>
<DIV>Gareth</DIV>
<DIV> </DIV>
<DIV><FONT face=System color=#800080 size=2>Ms. Gareth B. Davis</FONT></DIV>
<DIV><FONT face=System color=#800080 size=2><STRONG>Research Assistant
II</STRONG></FONT></DIV>
<DIV><FONT face=System color=#800080 size=2>Neuro-magnetics Division of
</FONT></DIV>
<DIV><FONT face=System color=#800080 size=2>the Department of
Neurology</FONT></DIV>
<DIV><FONT face=System color=#800080 size=2>Vanderbilt University Medical
Center</FONT></DIV>
<DIV><FONT face=System color=#800080 size=2>615-936-3318</FONT></DIV>
<DIV><FONT face=System color=#800080 size=2></FONT> </DIV>
<DIV> </DIV>
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<DIV><FONT size=2>-----Original Message----- <BR><B>From:</B> Rowani
[mailto:rowani.mohdrawi@student.adelaide.edu.au] <BR><B>Sent:</B> Mon
9/15/2003 12:08 AM <BR><B>To:</B> histonet@pathology.swmed.edu <BR><B>Cc:</B>
<BR><B>Subject:</B> [Histonet] Mice embryonic brain - EM Poor tissue quality -
need help<BR><BR></FONT></DIV>
<P><FONT size=2>Hi<BR>My electron microscopy sections of brain of mice
foetuses (18.5 dpc) are<BR>difficult to interpret because of
disrupted and lysed cells. The connective<BR>tissue surrounding the
cells appears disintegrated with empty<BR>spaces/vacuoles like artifact
?. The brain was processed according to the<BR>following protocols:
brain was dissected and immersed in ice cold fixative<BR>(glutaryldehyde 2.5%,
paraformaldehyde 4%, sucrose 2% in PBS pH 7.2) for 1-2<BR>hrs to firm it up
before cutting them to smaller fragments (2 cumm). After<BR>an overnight
fixation, re-fixed in osmium tetroxide, washed in PBS,<BR>dehydrated through a
series of ethanol (70%, 90%, 95% and 100%) and embedded<BR>in araldyte
resin. The brain of adult mice processed using similiar<BR>protocol was
excellent. COuld someone suggest ways to improve the tissue<BR>quality
?<BR><BR>TQ<BR><BR>Rowani<BR><BR><BR><BR><BR>_______________________________________________<BR>Histonet
mailing list<BR>Histonet@lists.utsouthwestern.edu<BR><A
href="http://lists.utsouthwestern.edu/mailman/listinfo/histonet">http://lists.utsouthwestern.edu/mailman/listinfo/histonet</A><BR></FONT></P></BLOCKQUOTE>
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