Ax10scopea1 microscope

Histological sections as well as hematoxylin and eosin staining was performed by the Utah Veterinary Diagnostic Laboratory using standard methods. Grayscale images were taken on a AX10 scope.A1 microscope (Zeiss) equipped with a computer-controlled charge coupled device (CCD) black-and-white camera (Zeiss Axiocam).

Immunohistochemistry was performed as described previously11 (link). Briefly, the mice were perfused with 4% formaldehyde fixative through the circulatory system, and postfixed with phosphate-buffered 4% formaldehyde and brain sections were cut at 60-μm thickness and every sixth section was stained. The antibody retrieval was performed at 80 °C for 20 min with Target Retrieval Solution (Dako). The sections were treated with methanolic H₂O₂ (3% H₂O₂ and 1% absolute methanol) for 30 min to inactivate endogenous peroxidase activity. The sections were then blocked with 0.5% bovine serum albumin (BSA) in TBS for 1 h at room temperature (RT), and incubated overnight at 4 °C with rabbit polyclonal anti-HSV-1 (1:500; DakoCytomation, B0116) antibody in tris-buffered saline (TBS) containing 0.1% BSA, 0.3% Triton-X, followed by several washes in TBS containing 0.3% Triton-X TBS and staining with and the secondary antibody, donkey anti-rabbit HRP (1:500; Sigma-Aldrich) for 1 h at RT. Horseradish peroxidase-labelled antibodies were developed with chromagen diaminobenzidine. Finally, the slides were rinsed in water, dehydrated and prepared for imaging. The imaging was performed on Zeiss AX10, Scope A1 microscope using × 20 and × 2.5 objectives.