[Histonet] fixed and unfixed immuno and antuibody oreps

From:"George Cole"

I promise to drop this line---I have had an itch to find out
something--- I realize I must give it up---I have asked it repeatedly
and I don't want to keep asking the same thing---impatience is bound to
develop.  Retired as I am---I no longer have lab in which to answer such
questions. Most histotechs nowadays are brought up doing one side of the
question---indeed it is the standard procedure and my question  just
doesn't scan with  today's techs.  But if I could shuck 10 years and be
back in my lab and I was given the responsibility to do a certain class
of work I would do the following:
Antibody technique or immunofluorescence procedure---any procedure that
has a fixative in its beginning followed by some procedure to neutralize
the effects of the fixative----I would run at least 3 trials on the 2
sets of pairs: 1) Tissue unfixed 2) tissue fixed, then processed with by
whatever step was supposed to neutralize the fixation effect, rinsing
the tissues carefully afterwards.  . Then I would run tissues 1 and 2 in
the same preparation.  And I would choose as my standard procedure which
ever procedure of the pair which gave the better results. I drop that
and run.  The crowding effect of Standard Procedure Itis is out there.
But then I might just find my question was trivial and standard
procedure might win.  But has the fix-unfix procedure ever been tested
against the unfixed prep? This retired picky-compulsive tech would never
run ANY procedure as standard without testing its
effectiveness---comparing it--- with any reasonable alternative
procedure for quality. Nuff said.  Retired Tech now enters a stage of
quiet upon the subject on the histonet, but I will still brood over the
untapped quality test in ANY procedure done by rote rather than via the
search for the most excellent way to do that test. 
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