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<DIV><FONT face=Arial size=2>George Cole</FONT></DIV>
<DIV><FONT face=Arial size=2>The slide was left out exposed to sunlight for a
prolonged period, probably several weeks and yes all our slides are coverslipped
using the same coverslipping tape.</FONT></DIV>
<DIV>.....................................................<BR>Peter
Smale<BR>Anatomical Pathology<BR>National Health Laboratory Service<BR>Chris
Hani Baragwanath Hospital<BR>Soweto, Johannesburg<BR>tel: +2711
4898711<BR>fax: +2711 4898717 <BR>e-mail: <A
href="mailto:peter.smale@nhls.ac.za">peter.smale@nhls.ac.za</A></DIV>
<DIV> </DIV>
<DIV>This message is for the designated recipient only and may contain
priviliged,<BR>prorietary, or otherwise privare information. If you have
received it in error,<BR>please notify the sender immediately and delete the
original. <BR>Any other use of the email by you is
prohibited.<BR>-----------------------------------------------------<BR></DIV>
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style="PADDING-RIGHT: 0px; PADDING-LEFT: 5px; MARGIN-LEFT: 5px; BORDER-LEFT: #000000 2px solid; MARGIN-RIGHT: 0px">
<DIV style="FONT: 10pt arial">----- Original Message ----- </DIV>
<DIV
style="BACKGROUND: #e4e4e4; FONT: 10pt arial; font-color: black"><B>From:</B>
<A title=georgecole@ev1.net href="mailto:georgecole@ev1.net">George Cole</A>
</DIV>
<DIV style="FONT: 10pt arial"><B>To:</B> <A title=peter.smale@nhls.ac.za
href="mailto:peter.smale@nhls.ac.za">'Peter Smale'</A> </DIV>
<DIV style="FONT: 10pt arial"><B>Sent:</B> Tuesday, November 25, 2003 10:43
AM</DIV>
<DIV style="FONT: 10pt arial"><B>Subject:</B> RE: [Histonet] restaining of
hematoxylin and eosin tissue sections</DIV>
<DIV><BR></DIV>
<DIV class=Section1>
<P class=MsoNormal><FONT face=Arial color=navy size=2><SPAN
style="FONT-SIZE: 10pt; COLOR: navy; FONT-FAMILY: Arial">Peter,<o:p></o:p></SPAN></FONT></P>
<P class=MsoNormal><FONT face=Arial color=navy size=2><SPAN
style="FONT-SIZE: 10pt; COLOR: navy; FONT-FAMILY: Arial">No clear answers from
me, but one vague guess<SPAN class=GramE>----<SPAN
style="mso-spacerun: yes"> </SPAN>What</SPAN> caused the H & E to
fade? H & E’s seem to hold up pretty well. Did a lot of people look at the
slide for prolonged periods under bright light? If not<SPAN class=GramE>,
<SPAN style="mso-spacerun: yes"> </SPAN>I</SPAN> would consider the
covering method----that’s the only difference that I can see between that H
& E and a million other <SPAN class=SpellE>unfaded</SPAN> H & E’s. It
doesn’t seem likely that it involved the reagents you used to remove the cover
and the residual of the covering, because the fading had already taken place.
Do you have any other H & E’s covered the same way?<SPAN
style="mso-spacerun: yes"> </SPAN>How are they? I hope you solve your
problem.<o:p></o:p></SPAN></FONT></P>
<P class=MsoNormal><FONT face=Arial color=navy size=2><SPAN
style="FONT-SIZE: 10pt; COLOR: navy; FONT-FAMILY: Arial">georgecole@ev1.net
<SPAN style="mso-spacerun: yes"> </SPAN><SPAN
style="mso-spacerun: yes"> </SPAN><SPAN
style="mso-spacerun: yes"> </SPAN>.<SPAN style="mso-spacerun: yes">
</SPAN><SPAN style="mso-spacerun: yes"> </SPAN><SPAN
style="mso-spacerun: yes"> </SPAN><SPAN
style="mso-spacerun: yes"> </SPAN><o:p></o:p></SPAN></FONT></P>
<P class=MsoNormal style="MARGIN-LEFT: 0.5in"><FONT face=Tahoma size=2><SPAN
style="FONT-SIZE: 10pt; FONT-FAMILY: Tahoma">-----Original
Message-----<BR><B><SPAN style="FONT-WEIGHT: bold">From:</SPAN></B>
histonet-admin@lists.utsouthwestern.edu
[mailto:histonet-admin@lists.utsouthwestern.edu] <B><SPAN
style="FONT-WEIGHT: bold">On Behalf Of </SPAN></B>Peter Smale<BR><B><SPAN
style="FONT-WEIGHT: bold">Sent:</SPAN></B> </SPAN></FONT><st1:date Year="2003"
Day="25" Month="11"><FONT face=Tahoma size=2><SPAN
style="FONT-SIZE: 10pt; FONT-FAMILY: Tahoma">Tuesday, November 25,
2003</SPAN></FONT></st1:date><FONT face=Tahoma size=2><SPAN
style="FONT-SIZE: 10pt; FONT-FAMILY: Tahoma"> </SPAN></FONT><st1:time
Minute="17" Hour="0"><FONT face=Tahoma size=2><SPAN
style="FONT-SIZE: 10pt; FONT-FAMILY: Tahoma">12:17
AM</SPAN></FONT></st1:time><FONT face=Tahoma size=2><SPAN
style="FONT-SIZE: 10pt; FONT-FAMILY: Tahoma"><BR><B><SPAN
style="FONT-WEIGHT: bold">To:</SPAN></B>
histonet@lists.utsouthwestern.edu<BR><B><SPAN
style="FONT-WEIGHT: bold">Subject:</SPAN></B> [Histonet] restaining of
hematoxylin and eosin tissue sections<BR><B><SPAN
style="FONT-WEIGHT: bold">Importance:</SPAN></B> High</SPAN></FONT></P>
<P class=MsoNormal style="MARGIN-LEFT: 0.5in"><FONT face="Times New Roman"
size=3><SPAN style="FONT-SIZE: 12pt"><o:p> </o:p></SPAN></FONT></P>
<DIV>
<P class=MsoNormal style="MARGIN-LEFT: 0.5in"><FONT face=Arial size=2><SPAN
style="FONT-SIZE: 10pt; FONT-FAMILY: Arial">I wonder if someone can assist me
in solving this problem.I retrieved a hematoxylin and eosin/phloxine stained
slide from 1998 and discovered it was faded.The tissue section was mounted in
a coverslipping machine that uses coverslipping tape composed of cellulose
triacetate and is coated on one side with resinous mountant.This is the
procedure I followed to decoverslip,decolorise and restain: I placed the slide
in acetone for 7 minutes to remove the coverslip and then put the slide
in xylene for 6 hours to remove any residual resinous mountant.I took the
slide through graded ethanols to water after which I decolorised with
acid/alcohol for one minute, took the slide back to water and restained the
nuclei with Gill's Hematoxylin ( for 5 min. ) and
counterstained with a mixture of 2% eosin y and 1% phloxine B with 4
drops of conc. glacial acetic acid added ( for 3 min. ).The nuclear staining
turned out pale and the counter stain was patchy almost as if something was
blocking the tissue's ability to retain the dyes I cannot figure out why the
tissue is not staining with the same intensity as it did in 1998.Please keep
in mind the slide was left out in natural light for a while which resulted in
it becoming faded.</SPAN></FONT><o:p></o:p></P></DIV>
<DIV>
<P class=MsoNormal style="MARGIN-LEFT: 0.5in"><FONT face=Arial size=2><SPAN
style="FONT-SIZE: 10pt; FONT-FAMILY: Arial">.....................................................<BR>Peter
Smale<BR>Anatomical Pathology<BR>National Health Laboratory Service<BR>Chris
Hani Baragwanath Hospital<BR>Soweto, Johannesburg, South
Africa<BR>tel: +2711 4898711<BR>fax: +2711 4898717
<BR>e-mail: <A
href="mailto:peter.smale@nhls.ac.za">peter.smale@nhls.ac.za</A></SPAN></FONT><o:p></o:p></P></DIV>
<DIV>
<P class=MsoNormal style="MARGIN-LEFT: 0.5in"><FONT face="Times New Roman"
size=3><SPAN style="FONT-SIZE: 12pt"> <o:p></o:p></SPAN></FONT></P></DIV>
<DIV>
<P class=MsoNormal style="MARGIN-LEFT: 0.5in"><FONT face=Arial size=2><SPAN
style="FONT-SIZE: 10pt; FONT-FAMILY: Arial">This message is for the designated
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information. If you have received it in error,<BR>please notify the sender
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prohibited.<BR>-----------------------------------------------------</SPAN></FONT><o:p></o:p></P></DIV></DIV></BLOCKQUOTE></BODY></HTML>