Anti hexon fitc conjugated antibody

Brain tissue was harvested and fixed in 4% paraformaldehyde for 72–96 hours. Tissues were paraffin embedded and sectioned at 10 μm in thickness, and hematoxylin and eosin (H&E) and Prussian blue staining was done every 10^th section to visualize the tumor and to detect ferumoxytol-labeled NSCs. Prussian blue staining was performed using the Accustain iron stain kit (Sigma-Aldrich, St. Louis, MO, USA). Images were captured with a 10× lens using Nikon-Eclipse 2E200U. Viral hexon protein staining was done using an anti-hexon FITC-conjugated antibody (Millipore, Billerica, MA, USA). In brief, sections were blocked using 1% BSA + 0.3% Triton-X for 1 h and then incubated with the antibody diluted in 1% BSA overnight at 4 °C. Sections were then washed in PBS and mounted using ProLong® Gold Antifade Mountant with DAPI (Life Technologies, Waltham, MA, USA). Imaging was performed at the University of Chicago Integrated Light Microscopy Facility. Images were captured with a Zeiss Axiovert 200m inverted epifluorescence microscope (Carl Zeiss Microscopy, Thornwood, NY, USA) with a Hamamatsu Orca ER CCD camera (Hamamatsu Photonics, Skokie, IL, USA) for fluorescence imaging run by SlideBook 5.5 software (Intelligent Imaging Innovations, Denver, CO, USA).