[Neuroscience] Densitometric immunoreactivity analysis

NMF nfournier at sasktel.net
Thu Dec 8 01:56:36 EST 2005


I am interested if anyone has prior experience performing densitometric 
analysis on rat brain sections using NIH Image or Image J software.  If so, 
what methods do you use to calibrate the software prior to taking readings? 
In addition, do you typically remeasure luminosity after every, say 12th 
image?  If so, how do you perform this measurement?   Secondly, I am 
interested in evaluating if there are specific changes confined to the 
different laminar layers of the hippocampus (i.e. CA3 stratum oriens, 
stratum radiatum, stratum lucidum; CA1 stratum oriens, stratum radiatum, 
etc.)   In order to quantify these specific areas, I thought the best method 
would be to drop numerous open square cursor boxes of a specific size 
(perhaps 40 pixel by 40 pixel) along the region of interest.  (I thought 
this would be a better alternative to direct tracing).   I am uncertain if 
NIH image allows you to set the size of the boxes, or if you can place 
multiple boxes on a captured image.  Does anyone know if this possible or 
can suggest a better way of obtaining this information?

I would appreciate any insights, solutions, and suggestions.


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