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Optimizing the Microfil Dye Perfusion Technique will Allow for Rat Cerebral Vascular Network Analysis through uCT Image Quantification
Author(s) -
McCarter Monika,
Balaez Alexander,
Yarrow Joshua,
Zubcevic Jasenka,
Raizada Mohan
Publication year - 2015
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.29.1_supplement.647.1
Subject(s) - perfusion , medicine , cerebral blood flow , heparin , perfusion scanning , cerebral perfusion pressure , anesthesia , cardiology
Vascular pathophysiology is a hallmark of hypertension (HTN)‐linked cardiovascular disease, as increased peripheral resistance leads to vascular remodeling. Consistent with this is the evidence of links between impaired cerebral blood flow and hypoperfusion of the brainstem in HTN (1‐2). Thus, establishment of a technique for the cast dye perfusion would allow for 3D quantification of the microvasculature in order to investigate cerebral vascular changes associated with HTN and stroke. Methods Male SHR and Wistar Kyoto (WKY) rats (3 and 12 week) were intracardially perfused using varying procedures. Focusing on limiting vessel damage and bubbling, Protocol 1 was as follows: 20 ml Microfil dye (47% dye, 51% diluent, 1% curing agent), 0.1 ml/min. Protocols 2,3,4 and 5 followed an identical procedure, however, perfusion speeds varied from 0.1 to 35 ml/min. A PBS/Nitroprusside/Heparin mixture was added to Protocol 6 to induce greater vessel dilation, prior to perfusion with a 1:4 dye to diluent ratio, with 0.2 ml of curing agent and 0.5 ml heparin at 50 ml/min. All procedures allowed for curing overnight at 4C. Results uCT imaging revealed that Protocol 6 provided the best perfusion with minimum damage to microvessels. Conclusion Thus, our study has established a protocol to generate 3D images of the brain vasculature and puts us in a unique position to compare the impact of HTN on microvessels in the autonomic brain regions. Acknowledgements This research was made possible through funding from the APS UGSRF program. Special thanks to Mohan Raizada and lab for all of their support and guidance.