Journal: Bio-protocol
Article Title: Utilizing EdU to Track Leukocyte Recruitment to the Brain
doi: 10.21769/BioProtoc.5527
Figure Lengend Snippet: EdU was injected i.p. one day before subjecting the mouse to transient middle cerebral artery occlusion (tMCAO) surgery. After 24 h, the mouse was euthanized and the brain recovered, sliced into 1 mm sections, fixed, embedded in optimal cutting temperature (OCT) media, and snap frozen on dry ice. Coronal slices (25 μm) were acquired with a cryostat and mounted on Superfrost Plus slides. Tissue was processed for EdU detection as described above and imaged using widefield microscopy at 10× magnification (A). Insets show individual channels for EdU (B, cyan), tdTomato (C, red), and vessels (D, green). Arrows indicate EdU-positive neutrophils, and open arrowheads indicate EdU-negative neutrophils. Widefield image acquisition on 1 or 2 mm sections is straightforward, making it possible to capture the entire brain cross-sections at 10× or even 20×. These image series can be stitched together using a number of free or commercial software programs. We prefer NIS Elements and FIJI. All scale bars are 50 µm.
Article Snippet: NIS Elements Advanced Research Imaging Software (Nikon, version 5.41.02) 2.
Techniques: Injection, Microscopy, Software