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. 2016 Aug 3;7:330. doi: 10.3389/fphys.2016.00330

Figure 2.

Figure 2

Characterization of Hb following preterm rabbit pup IVH. The presence of Hb following IVH was characterized within the brain utilizing the inherent peroxidase activity of Hb. Rabbit pups with IVH or sham controls were euthanized at 72 h of age followed by saline and freshly prepared 4% PFA perfusion. Afterwards the brains were dissected out from the skulls and immersed in 4% PFA. Brains were prepared and sections and a number of subventricular and periventricular anatomically comparable regions of interests (ROI) were chosen. Neuroanatomically defined ROI's located in the rostral forebrain at the levels of rostral forebrain (Level 1), caudal forebrain (Level 2), rostral midbrain (Level 3) and caudal midbrain (Level 4), were stained for peroxidase activity of Hb as described in the Materials and Methods Section. 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 40 × magnification lens. Images used for illustrations, from regions of interest (ROI), were grabbed with the viewer software NDP.view2 Viewing software. Scale bar of slide scan image indicate 2.5 mm and of ROI images indicate 500 μm.