[Histonet] Fixed VS unfixed tissues for immunofluorescence and antibody work

From:"George Cole"

My histological career began in June 1962.  When I was assigned immuno
studies, the thinking of the day was that fixation blocked/ruined those
reactions as well as most antibody preps. SO it went until about 1980 or
so, when muscle and nerve biopsy work got so heavy, those immuno and
antibody needs were met in the main lab. Somewhere in there,  I got word
that immuno and antibody studies were being done on fixed tissues.  This
was startling news to me---a reversal of an Aboslute.  And I tell you,
such is the power the question-generating innards of my head, I would
have compared with-and-without-fixative preps of any immuno or antibody
studies that I might have been assigned. But I was hard enough pressed
to deal with my work load---I always picture biopsy patients , waiting
anxiously for  the results of their tissues. So the question of
fixed-unfixed immunos and antibody work didn't really revive until I
retired and became a regular visitor to the Histonet.  I know that I am
a born fuss budget and I would NEVER opt for ANY choice of  diagnostic
procedure without rating it with all candidate  procedures for that
specific study.  So far, I have had no comforting word from the
world-wide Histology community that EVERY immuno and antibody technique
now being discussed on the Histonet had been cleared for action by
initial comparisons of results with and without fixation. All I have
seen in mentioning this picky-picky compulsion of mine, is the flat
statement is THIS IS HOW WE DO IT!! The  work being done in any lab is
almost never reported with credentials of the procedures used---that
that particular procedure was being done, because it had  won a contest
of effectiveness over other procedures.  Furthermore, just beneath the
surface, is the feeling that the "This is the way I do it. Period!!"
attitude was lurking. Now, this aging retiree wishes to avoid conflict
and arousing tempers in you younguns. Age does not cater to warfare.
But I would treasure any reports from you active histotechs about the
credentials of choice from among candidate procedures--- the winners of
effectiveness resulting from  comparisons of techniques that are rivals
to your chosen ones.  Such reports would receive an honored place,
hanging on the wall here over my computer.  I would honor them---and you
would add peace to the Age-Naps of this old histotech!!.
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