Nuance fx multiplex biomarker imaging system

Modified Perl's histochemistry was performed on 14‐μm cryostat sagittal sections of 4% paraformaldehyde‐fixed brain. After a preliminary incubation with 2% ammonium sulfide, sections were incubated with 10% potassium ferricyanide and 0.5% HCl, followed by incubations with 0.025% diaminobenzidine and 0.005% H₂O₂, and counterstained with 0.5% Cresyl violet. Staining at different emission wavelength was evaluated by the Nuance^® FX multiplex biomarker imaging system (PerkinElmer) in 10 different fields for each mice. A semi‐quantitative analysis of iron deposition normalized for cell staining in areas of defined size was performed using ImageJ software.

Modified Perl’s staining was performed on 14 μm sagittal sections of paraformaldehyde fixed brain as described in^{20 (link)}. Staining at different emission wavelength was evaluated by the Nuance® FX multiplex biomarker imaging system (PerkinElmer) in the choroid plexus of the mice. A semi-quantitative analysis of iron deposition normalized for cell staining was performed using ImageJ software (National Institute of Health, http://rsb.info.nih.gov/ij) as in ref. ^{20 (link)}.