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.