[Histonet] section thickness

[Rittman, Barry R]

This is a difficult question to answer as it depends on exactly what you
wish to measure.

In general
The thinner the sections the more precisely you can locate and measure
the reaction product but the fewer entire cells will be seen. 
If you are counting cells then the thicker the sections and the more
localized the reaction product the better as otherwise you should apply
a correction factor (as for example per Abercrombie 1946). This is
because in image analysis of thinner sections it is often difficult to
set limits to exclude partial cells.

Another point to consider is that of determining the section thickness.
Most assume that if you cut two blocks at 4 micron section thickness
that you will be able to compare the sections for density of reaction
product. However we all know that it is difficult to get all sections of
the same exact thickness. If one is at 4 microns and the next at 3
microns then the second has 25% less volume of tissue and presumably
will have less total reaction product.  There are steps that can be
taken to minimize or eliminate this error such as including a material
of know optical density in the blocks that are cut along with the
tissue.
Hope that these comments are useful.

Barry


-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu
[mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Edwards,
R.E.
Sent: Thursday, May 18, 2006 5:44 AM
To: histonet <@t> pathology.swmed.edu
Subject: [Histonet] section thickness

 
 
 
In your  invaluable, but,  no  doubt differing opinions, what  is  the
optimal section thickness   for formalin fixed paraffin processed
tissues for  immunostaining, using  DAB  as  chromogen?, there has been
some  discussion here  whether  it  should be 4,5,6, or  7microns, many
thanks.
 
 
Richard  Edwards
 
 
MRC TOXICOLOGY UNIT
 
LEICESTER..U.K.......
 
 
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