Re: Trying to measure nuclear volume using IMAGEJ

Posted by Gabriel Landini on Feb 17, 2009; 9:38pm
URL: http://imagej.273.s1.nabble.com/Trying-to-measure-nuclear-volume-using-IMAGEJ-tp3693666p3693669.html

On Tuesday 17 February 2009 21:07:14 Doube, Michael wrote:
> I tried Feulgen staining on tendon some years ago and it was pretty harsh
> on the tissue; intercalating dyes like propidium iodide, Hoechst or DAPI
> may be kinder.  I can't speak for the quantitative, fluorimetric value of
> these stains though as I only used PI for nuclear morphology.

Some time ago Darek Kedra posted this reference, which is very good:
David C. Hardie, T. Ryan Gregory, and Paul D.N. Hebert, “From Pixels to
Picograms: A Beginners' Guide to Genome Quantification by Feulgen Image
Analysis Densitometry,” J. Histochem. Cytochem. 50, no. 6 (June 1, 2002):
735-750.

I however fail to understand why the authors suggest not to use background
correction (they do not give an explanation either). After all, if background
correction is not done, one cannot be sure of measuring the darkness of the
Feulgen stain or a dark spot on the illumination field.

Cheers

G.