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<p class=MsoNormal><span class=GramE><font size=2 face=Arial><span
style='font-size:10.0pt;font-family:Arial'>HISTOTECHS DOING IMMUNOFLUORESCENCE
SSUDIES.</span></font></span><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'>I just visited the hospital I retired from. In talking with
the great gal who replaced me and my good friend who runs the <span
class=SpellE>immuno</span> lab I mentioned that when I did <span class=SpellE>immunofluorescence</span>
studies on kidney, I cut sections thicker than anyone else---I was shaky <span
class=GramE>with <span style='mso-spacerun:yes'> </span>2</span> or 3 micron
sections as a beginner, so I cut one 3 micron, then went up to 5 microns,<span
style='mso-spacerun:yes'> </span>and, because I had been doing nothing
but muscles and nerves in which I did 8 and 10 micron sections, I included an 8
micron section. Well the kidney turned out to fluoresce with two preps. The 3
micron section was not very dramatic.<span style='mso-spacerun:yes'>
</span>The 8 micron section was brilliant, but a bit <span class=SpellE>globby</span>;
but to my surprise, all hands chose the 6 micron section.<span
style='mso-spacerun:yes'> </span>Also, as I had developed the multiple
sections techniques on muscles and nerves, I cut one 3 micron section for slide
1, 2 and 3, then went back and cut a 6 micron section for slide 1, 2 and 3, and
did the same for 8 microns sections. Well, I never went back to the 2 or 3
micron sections. I made 6 microns standard----and got many beautiful photos
brilliantly shining. The 8 micron section <span class=GramE>was <span
style='mso-spacerun:yes'> </span>a</span> little much, so I used that only
with bigger pieces of tissue. But remember what we are doing in this
procedure----we are NOT staining fine nuclear detail that takes thin sections
to do justice to---we are staining BLOBS and the photos of <span class=GramE>the
<span style='mso-spacerun:yes'> </span>6</span> microns sections use to be
passed around getting OOOHHED and AAAHHHED by the residents. Also, about the
multiple sections---this wasn’t always possible, because the kidney
biopsies are usually quite small and I had to leave tissue enough to do the
Jones silvers----but in this case, of the three sections <span class=GramE>on
one positive prep</span>, only one of the sections lit up. And what did that
tell me?<span style='mso-spacerun:yes'> </span>It told me that a single
section per slide work up on this case could miss being positive something like
66% of the time.<span style='mso-spacerun:yes'> </span>So I kept doing a
3 micron and as many 6 micron sections cut for each slide as the size of the
tissue would allow. The 3 micron section NEVER was positive, by chance or by
whatever----and I passed around many brilliantly lighted kidney sections photos
of the 6 micron sections.<o:p></o:p></span></font></p>
<p class=MsoNormal><font size=2 face=Arial><span style='font-size:10.0pt;
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><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><span
style='mso-spacerun:yes'> </span><o:p></o:p></span></font></p>
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