Ribbon scanning confocal for high-speed high-resolution volume imaging of brain
Fig 5
Multicolor large-volume imaging by ribbon scanning confocal microscopy.
A mouse was infected subcutaneously with VEEV TrD TaV-cherry (red) and at 96 hours post infection, fluorescent beads (green) were introduced into the vasculature by perfusion. The brain was harvested, sectioned approximately 4mm thick, and cleared by CUBIC. The section was imaged approximately two millimeters deep (the limit of the Olympus 25x, 1.05NA objective) on both sides and reconstructed as (A) one volume. (B) A single optical plane demonstrates that both the microvasculature and virus infected cells were observed throughout the volume. (C-D) Zoomed regions from a single optical place demonstrate that the vasculature in some areas with virus replication were inaccessible to fluorescent beads administered by perfusion.