Additional file 1 of Herpes simplex virus infection induces necroptosis of neurons and astrocytes in human fetal organotypic brain slice cultures

Additional file 1: Figure S1. Generation and metabolic activity of cultured human fetal organotypic brain slices. (A) Representative images showing unprocessed human fetal brain tissue and (B) selection of cortical human fetal brain tissue, as indicated by the white arrow, for subsequent slicing. (C) Agarose pieces were glued to the vibratome platform, as indicated by the white arrows. (D) Vertical and (E) horizontal orientation of the brain slice tissue dimension were employed to ensure standardization of the brain slice size. (F) Collection of the human fetal brain slice tissue in six separate collection chambers in artificial cerebrospinal fluid (aCSF) under constant oxygenation (95% O2, 5% CO2), as indicated by the white arrow. (G) Slicing of fetal brain slices was performed in aCSF under constant oxygenation using a vibratome. The slices were kept in an icocold environment to decrease cellular metabolic activity. (H) Viability assessment of a human fetal organotypic brain slices showing metabolic activity (purple) over a 14-days culture period using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. Images are representative of two independent subjects.