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<p class=MsoNormal><font size=2 face=Arial><span style='font-size:10.0pt;
font-family:Arial'>For Those Who Received Muscle & Nerve packets;<o:p></o:p></span></font></p>
<p class=MsoNormal><font size=2 face=Arial><span style='font-size:10.0pt;
font-family:Arial'>Sorry, when I made up the packets, I used forms filled out
for our last inspection by office staff.<span style='mso-spacerun:yes'>
</span>The typing was so much better than my own.<span
style='mso-spacerun:yes'> </span>All the ingredients were listed, but the
actual PAS procedure was left out.<o:p></o:p></span></font></p>
<p class=MsoNormal><font size=2 face=Arial><span style='font-size:10.0pt;
font-family:Arial'>The following procedure is done after the PAS reagents are
prepared:<o:p></o:p></span></font></p>
<p class=MsoNormal><font size=2 face=Arial><span style='font-size:10.0pt;
font-family:Arial'><span style='mso-spacerun:yes'> </span><o:p></o:p></span></font></p>
<p class=MsoNormal><font size=2 face=Arial><span style='font-size:10.0pt;
font-family:Arial'>This follows page 12:<o:p></o:p></span></font></p>
<p class=MsoNormal><font size=2 face=Arial><span style='font-size:10.0pt;
font-family:Arial'><o:p> </o:p></span></font></p>
<p class=MsoNormal><font size=2 face=Arial><span style='font-size:10.0pt;
font-family:Arial'>1. Put slides in the fixative<span
style='mso-spacerun:yes'> </span>10 minutes to one hour.<o:p></o:p></span></font></p>
<p class=MsoNormal><font size=2 face=Arial><span style='font-size:10.0pt;
font-family:Arial'>2. Rinse well in <span class=SpellE>deionized</span> water.<o:p></o:p></span></font></p>
<p class=MsoNormal><font size=2 face=Arial><span style='font-size:10.0pt;
font-family:Arial'>3. Place in freshly <span class=GramE>mixed<span
style='mso-spacerun:yes'> </span>0.5</span>% aqueous Periodic Acid<span
style='mso-spacerun:yes'> </span>5 minutes.<o:p></o:p></span></font></p>
<p class=MsoNormal><font size=2 face=Arial><span style='font-size:10.0pt;
font-family:Arial'>4. Wash thoroughly in <span class=SpellE>deionized</span>
water.<o:p></o:p></span></font></p>
<p class=MsoNormal><font size=2 face=Arial><span style='font-size:10.0pt;
font-family:Arial'>5. Place in <span class=GramE>Schiff’s Reagent</span><span
style='mso-spacerun:yes'> </span>15 minutes.<o:p></o:p></span></font></p>
<p class=MsoNormal><font size=2 face=Arial><span style='font-size:10.0pt;
font-family:Arial'>6. Rinse in three changes of acid rinse<span
style='mso-spacerun:yes'> </span>for two
minutes each.<o:p></o:p></span></font></p>
<p class=MsoNormal><font size=2 face=Arial><span style='font-size:10.0pt;
font-family:Arial'>7.<span style='mso-spacerun:yes'> </span>Wash in
gently running water<span style='mso-spacerun:yes'>
</span>for a minute or two.<span style='mso-spacerun:yes'> </span>Rinse
with <span class=SpellE>deionized</span> water.<span
style='mso-spacerun:yes'> </span>Stain with Gill’s <span
class=GramE>III<span style='mso-spacerun:yes'> </span><span class=SpellE>hematoxylin</span></span>
for <o:p></o:p></span></font></p>
<p class=MsoNormal><font size=2 face=Arial><span style='font-size:10.0pt;
font-family:Arial'><span style='mso-spacerun:yes'> </span>30 seconds to a
minute.<span style='mso-spacerun:yes'> </span>Blue and rinse with running
tap water<span style='mso-spacerun:yes'> </span>15 minutes.<span
style='mso-spacerun:yes'> </span>Rinse in <span class=SpellE>deionized</span>
water.<o:p></o:p></span></font></p>
<p class=MsoNormal><span class=GramE><font size=2 face=Arial><span
style='font-size:10.0pt;font-family:Arial'>8 .</span></font></span><font
size=2 face=Arial><span style='font-size:10.0pt;font-family:Arial'> Dehydrate
with 95% and 100% alcohols.<span style='mso-spacerun:yes'> </span>Clear
in <span class=SpellE>xylene</span> and mount in <span class=SpellE>Permount</span>
or equivalent.<o:p></o:p></span></font></p>
<p class=MsoNormal><font size=2 face=Arial><span style='font-size:10.0pt;
font-family:Arial'><o:p> </o:p></span></font></p>
<p class=MsoNormal><span class=GramE><font size=2 face=Arial><span
style='font-size:10.0pt;font-family:Arial'>DIASTASE<span
style='mso-spacerun:yes'> </span>DIGESTION</span></font></span><font
size=2 face=Arial><span style='font-size:10.0pt;font-family:Arial'><span
style='mso-spacerun:yes'> </span>PROCEDURE:<o:p></o:p></span></font></p>
<p class=MsoNormal><font size=2 face=Arial><span style='font-size:10.0pt;
font-family:Arial'><o:p> </o:p></span></font></p>
<p class=MsoNormal><font size=2 face=Arial><span style='font-size:10.0pt;
font-family:Arial'>1. Fix for 10 minutes.<o:p></o:p></span></font></p>
<p class=MsoNormal><font size=2 face=Arial><span style='font-size:10.0pt;
font-family:Arial'>2. Rinse well.<o:p></o:p></span></font></p>
<p class=MsoNormal><font size=2 face=Arial><span style='font-size:10.0pt;
font-family:Arial'>3. Digest in filtered Diastase solution for one hour at 37C.<o:p></o:p></span></font></p>
<p class=MsoNormal><font size=2 face=Arial><span style='font-size:10.0pt;
font-family:Arial'>4.<span style='mso-spacerun:yes'> </span>Rinse well in
<span class=SpellE>deionized</span> water.<o:p></o:p></span></font></p>
<p class=MsoNormal><font size=2 face=Arial><span style='font-size:10.0pt;
font-family:Arial'>5 Proceed with regular PAS routine from step 3 above.<o:p></o:p></span></font></p>
<p class=MsoNormal><font size=2 face=Arial><span style='font-size:10.0pt;
font-family:Arial'><span style='mso-spacerun:yes'> </span><o:p></o:p></span></font></p>
<p class=MsoNormal><font size=2 face=Arial><span style='font-size:10.0pt;
font-family:Arial'><span
style='mso-spacerun:yes'> </span><o:p></o:p></span></font></p>
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