Table 5 Detailed protocol for whole mice brain preparation for micro-CT with iodine.
From: High-resolution micro-CT for 3D infarct characterization and segmentation in mice stroke models
Step | Solution | Time (min) | Temp. (°C) |
|---|---|---|---|
Perfusion | |||
Lethal anesthesia | Ketamine/medetomidine, IP | 5 | 22 |
Blood perfusion | PBS | 10 | 22 |
Brain fixation | 4% PFA, in PBS | 10 | 22 |
Post-fixation | |||
Fixation | 4% PFA, in PBS | 7-60 days | 4 |
Staining | |||
Dehydration | 30%, 50%, 70%, 80%, 90% ethanol | 2 h, each | 22 |
Iodine | 1% iodine, in 90% metanol (15 mL/brain) | 24 h | 22 |
micro-CT prep | |||
Preparation | Parafilm® wrapping | 2 | 22 |
Brain Storing | |||
Short-term | Protected from light; Sealed container | 3–12 months | 4 |
Long-term | Dehydration; Resin embedding | Years | 22 |
