Fig. 3.

Immunofluorescence labeling of Hb and the i.c.v. administered rA1M. Images represent the double IF-labeled Hb (red) and A1M (green) together with the nuclear staining DAPI (blue), in animals with IVH that were treated with rA1M (IVH + rA1M). Selected ROIs of peroxidase activity from corresponding areas, recorded in parallel sections. Rabbit pups were euthanized at 72 h of age followed by saline and freshly prepared 4% PFA perfusion. Afterwards brains were prepared and sections at the levels of rostral forebrain (Level 1) and rostral midbrain (Level 3), were immunolabeled for Hb (red) and A1M (green) as described in the “Materials and methods” section. Fluorescence microscope analyses were performed on a wide-field Olympus microscope (IX73) and slide scanning were performed on a Hamamatsu NanoZoomer 2.0-HT Digital slide scanner: C10730. Scanning was performed with a 40x magnification lens. Images used for illustrations, from ROIs, were grabbed with the viewer software NDP.view2 Viewing software. Scale bar of slide scan images indicates 2.5 mm and in ROI images 25 μm