RE: Quality of Fixation

From:Linda Jenkins

Charles, that explains it!  Well... yes... that could be a 
problem.  Ours is such a qualitative science it can be frustrating 
sometimes.  How about Gas Chromatography(GC)?  You could take a portion of 
fresh unfixed tissue, your perfused section, and a portion from the 
un-improved perfusion sample. Do you have the availability of GC?  If not, 
a university will usually have one.
         Good Luck!
P.S.  I will try to load the picture today...thanks for the info.

  At 09:40 PM 3/9/2003 -0600, you wrote:
Thanks, to load a picture on the web for histonetters to see, give it a
name and send it to  They will post it so anyone coming to will can see it, then you reference it by name in an email
to the histonet at the usual address.

I claim a better way to perfuse over other ways to perfuse (no
shrinkage, no red blood cells left), the grant reviewer says show that
the fixation is at least as good.  I see it is, but what quantitative or
visual measure (intensity of staining, evenness of staining, etc. can I
use to prove it?  A measure sensitive only to fixation?

So far, the answer is there is not standard test to quantify degree of
fixation.  Some interesting suggestions about proteins stains that are
sensitive to degree of fixation.  I know from experience that HRP
reactions do not work on unfixed tissue, but also that overfixation
reduces HRP sensitivity.


Charles W.  Scouten, Ph.D.
5918 Evergreen Blvd.
St. Louis, MO 63134
Ph: 314 522 0300
FAX  314 522 0377

Linda Jenkins, HT
Clemson University
Dept. of Bioengineering
Clemson, SC 29634-0905

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