Heart ink injection
Lab/Group: Cossu Lab
Related Journal & Article Information
Journal: Nature Medicine
Article Title: PlGF–MMP-9–expressing cells restore microcirculation and efficacy of cell therapy in aged dystrophic muscle
Introduction
With this technique it is possible to label the blood vessel in vivo in order to analyze in whole mount the quantity and quality of blood vessel.
Materials
Reagents
1,0ml syringe
China Black Ink
Stadard anesthetic (Ketamine and Xylazine)
Equipment
Procedure
1. Anesthetize the mouse
2. Using a 1 ml syringe withdraw 0,5 ml of mouse blood from the heart ventricle
3. Inject 1ml of China Ink and wait for 30 sec (until the eyes, mouth and legs became black)
4. Quickly dissect and fix in 4% PFA over night at 4°C
5. Treat the sample with a solution of 0,5% KOH for one day
6. Treat the sample with a solution of 0,5% KOH/Glycarol 2:1 one day
7. Treat the sample with a solution of 0,5% KOH/Glycerol 1:1 for one day
8. Treat the sample with 100% Glycerol for one day (in order to visualize clearly all the inner blood vessels)
Troubleshooting
Critical Steps
Injection in the heart after blood withdrawal
Anticipated Results
eyes, mouth and legs black stained
References
Acknowledgements
Keywords
Heart, inke injection, blood vessel
