We use Prussian Blue staining of mouse brain sections from a model of inherited iron overload and it works well. I wouldn't use it as a quantitative measure but rather a qualitative one. There is a lower limit of detection of course below which you will not see any stain even though there is iron present in the tissue. You can also use DAB enhancement to make the stain more sensitive although this can also increase background staining.

Title: Iron accumulation in Alzheimer disease is a source of redox-generated free radicals

Author(s): SMITH, MA; HARRIS, PLR; SAYRE, LM; et al.

Source: PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA Volume: 94 Issue: 18 Pages: 9866-9868 Published: SEP 2 1997

Dear Anand,

not knowing whether you can work with modern approaches like the mentioned PIXE-method (by Markus Morawski above) I would like to point you to (classical histochemical approach) an

,

published by

Roschzttardtz H, Conéjéro G, Curie C and Mari St.

Published online before print September 2009, doi: http://dx.doi.org/10.1104/pp.109.144444

in Plant Physiology November 2009 vol. 151 no. 3 1329-1338

available at: http://www.plantphysiol.org/content/151/3/1329.full.

< The result confirms that the sensitivity of the Perls method is drastically enhanced by the DAB/H2O2 treatment in plant tissues > (not knwoing whether the article is free download or PPV= pay per view, in latter case you can write to me....).

Best wishes and good luck,

Wolfgang

@Morawski : Thank you for the response. I do agree PIXE is reliable, but sadly we don't have access to the facility. We have to resort to simpler methods.

@Muss: Thanks sir for the response. I'll definitely take a look at the work you pointed out.

Dear Anand,

your'e welcome. I thought it might be helpful also to include more and "classical" hsitochemical approaches too since neither I (sadly) have the possibility to do PIXE-analyses. Can you access the mentioned article?

Thank you for your kind reply. all my best wishes and regards, Wolfgang

Thanks Drs. Perry and Muss. I have access to the article. I shall go through and discuss with our histologist.

We use Prussian Blue staining of mouse brain sections from a model of inherited iron overload and it works well. I wouldn't use it as a quantitative measure but rather a qualitative one. There is a lower limit of detection of course below which you will not see any stain even though there is iron present in the tissue. You can also use DAB enhancement to make the stain more sensitive although this can also increase background staining.

Thank you so much for your response.

I have studied hemochromatosis (hemosiderosis is a better term, because hemochromatosis is a sytemic disease,  with iron deposits absent in the brain) in the human brain for a long time. I found Turnbull (ferrocyanid after pretreatment with (smelly) H2S) more sensitive than Perls (ferricyanid), because HCl, as pretreatment for Perls, dissolves part of the iron. But remember that neither of the methods shows "organic" iron (negative on hemoglobin, for instance, and  on iron-rich brain nuclei). 

Perls staining is pretty effective, easey and low cost.  We used it here.  The colour contrast is pretty good for image analysis.

It works very well. You can also enhance it with DAB to increase sensitivity

In short I point out to the possible use of Staining Perl's Prussian Blue reaction for ferriciron positivity in capillary endothelial cells of the blood-brain barrier.I hope that the my and coauthors published work can give you an contribution and help in your planed study.

Above-mentioned article you can find on the link:

http://dx.doi.org/10.4236/abb.2014.51003

Late reply to Lucijan:

since I found this your reply #15 just some minutes ago I tried to access the Article you mentioned in your RE#015 above. For convenience only I would like to add here the bibliographical data:

"§Above-mentioned article you can find on the link:

http://dx.doi.org/10.4236/abb.2014.51003 " ==>

NB: links to: http://file.scirp.org/Html/3-7300811_41659.htm (or - http://file.scirp.org/Html/3-7300811_41659.htm#txtF2

cf: automatic conversion into the RG-database-title: Article Methemoglobinemia-A biomarker and a link to ferric iron accu...

= Lucijan Mohorovic, et al. 2015.

Advances in Bioscience and Biotechnology Vol.5 No.1(2014), Article ID:41659,7 pagesDOI:10.4236/abb.2014.51003

Methemoglobinemia—A biomarker and a link to ferric iron accumulation in Alzheimer’s disease

Since in that Article there is no real mention / explanation of the Perl's reaction technique in the 'Methods'-section (cf.: "2.Methods and Results"),

I would like to ask ( or perhaps give already here the right information )

whether it is correct you used the staining technique as mentioned/referenced in your earlier, interesting and 'well-done', excellent article:

Article Brain iron accumulation in unexplained fetal and infant deat...

Brain iron accumulation in unexplained fetal and infant death victims with smoker mothers-The possible involvement of maternal methemoglobinemia

by LAVEZZI et al, BMC Pediatrics_2011_11:62 ==> OPEN ACCESS at:

http__s://bmcpediatr.biomedcentral.com/articles/10.1186/1471-2431-11-62; NB: to access the journal's pages (OPEN ACCESS), copy the latter URL but - prior to pasting into your browser, delete the two underline signs "__" in between >http __ and s