Visualization of murine in situ neurovascular interfaces
Step-by-step bench protocol — companion to Cell Reports Methods (2021)
Read original publicationThis STAR Protocol is the step-by-step lab guide for the iterative micro-CT workflow validated in Cell Reports Methods (2021). Use this page for bench execution — specimen selection, reagent guidance, and quality checkpoints. The shared workflow diagram and Imaging Suite registration steps below apply to both publications.
Video walkthroughs for this workflow are on the methods paper overview.
Sample prep
Sample prep
Bench protocol for specimen preparation — follow STAR timing, reagent specifications, and quality checkpoints before the first scan.
- Select age and genetic background appropriate to the experimental question.
- Retrograde aortic or transcardial perfusion, fixation, and cranial integrity checks per protocol.
- Infuse low-density polymer mixture with confirmation of arterial and venous filling.
- Document reagent batches, processing times, and QC checkpoints for multi-lab reproducibility.
Shared workflow
One specimen · multiple scans
- Vascular casting
- Scan I · bone + vessels
- Decalcify
- Scan II · vessels
- PTA stain
- Scan III · soft tissues
Iterative imaging
Acquire multiple micro-CT volumes from the same intact specimen — each processing stage reveals a distinct anatomical layer without sectioning.
- Scan I captures bone and cast vasculature together.
- Controlled decalcification removes bone mineral; Scan II captures the isolated vascular cast.
- PTA staining followed by Scan III adds contrast-enhanced soft tissues for full multimodal context.
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