Kim Annink

54 Chapter 3 Figure 1: The template for ADC measurements at the level of the pons and vermis that was copied to all scans and slightly adapted per patient. Postmortem analyses The histopathological examination of the brains of the infants with HIE was performed in the UMC Utrecht using the following protocol: sections of various brain regions, including cerebellum and vermis, were obtained after removing and fixing the brains in 4% buffered formalin for three to four weeks. After fixation, a transverse section of the vermis and central section of the cerebellar hemisphere with DN were sampled and embedded in paraffin. Histological sections of 5-6 μm thick were cut and mounted on coated slides. Slides were stained with Hematoxylin and Eosin (H&E) or with monoclonal antibodies against CD68 (Novo Castra, Newcastle, UK) and HLA-DR (Dako, Glostrup, Denmark). The H&E staining was used to study the morphology and number of PC’s. The CD68 and HLA-DR stains were used to visualize microglia and macrophages activation (13). The tissue was