Eight weeks after the AV loop implantation, animals were anesthetized with isoflurane and the vascular system was perfused with the radiopaque agent Microfil® MV-122 (Flowtech Inc., Carver, MA, USA). The abdominal aorta was cannulated and V. cava inferior opened. Subsequently, the vascular system was first flushed with warm heparinized (100 I.E/mL) physiological sodium chloride solution (Ratiopharm GmbH, Ulm, Germany) until there was clear backflow and then flushed with 30 mL yellow Microfil® MV-122 containing 5% of MV Curing Agent (both from FlowTech Inc.). The perfused rats were stored at 4 °C for 24 h for hardening of the Microfil® solution. Subsequently, constructs were explanted and fixed in formalin for 24 h at 4 °C.
Mv curing agent
Manufactured by Flow Tech
Sourced in United States
The MV Curing Agent is a laboratory equipment product designed to facilitate the curing process. It functions as a catalyst, accelerating the chemical reactions involved in curing various materials. The MV Curing Agent is a essential tool for researchers and professionals working in materials science, chemical engineering, and related fields.
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4 protocols using mv curing agent
1
Vascular Perfusion and Tissue Fixation
2
Micro-CT Imaging of Vascular Networks
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For micro-computed tomography (micro-CT), 20 ml Microfil Silicone Rubber (MV-122, Flow Tech, Carver, MA, USA) containing 5% of MV Curing Agent (Flow Tech) was applied instead of India ink in one animal (POD 15). A customized Micro-CT scanner (Y.Fox, Yxlon, Garbsen, Germany) with an open multifocus X-ray tube (10–160 kV; focal spot sizes 1–5 μm), a CNC manipulator, and a 14-bit direct amorphous silicon flat panel detector (Varian PaxScan 2520 D/CL; Varian, Palo Alto, CA, USA) was used. Data were analyzed with Osirix v. 4.1.1 (Pixmeo, Geneva, Switzerland).
3
Lung Vascular Imaging via Microfil Perfusion
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Mice were anesthetized with avertin and euthanized by cutting off the abdominal aorta. The lung was perfused with 0.9% saline through the right ventricle, followed by perfusion using 5 mL of previously prepared Microfil (2.5 mL MV Compound +2.5 mL MV Diluent +4% MV-Curing Agent, Flow Tech Inc, MV-120). The lung was left for 30 min for the contrast agent to polymerize and was harvested and fixed in a 10% formalin solution. The micro vessels in lungs were imaged using a micro-CT SkyScan1276 (BRUKER), with settings as follows: 50 kV, 180 μA, pixel size 3.05 μm, 0.2-degree rotation step, and 180-degree rotation. The CT section images were captured using a CTAn software to reconstruct three-dimensional images.
4
Microvascular Imaging of Transplanted Muscle
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After two weeks of implantation, rats were again anesthetized as described above and placed in the supine position. The left hind limb was dissected and the isolation chamber was opened to expose the transplanted gastrocnemius muscle (Figure 5 A). The muscles were taken out of the chamber and stimulation was carried out as described above. After that, muscles were perfused with the contrast agent Microfil® as described previously [13 (link)]. Briefly, laparotomy incision was performed and animals were perfused by flushing the aorta with 150 mL of heparine solution and afterward with 20 mL yellow Microfil® (MV-122) containing 2.5% of MV Curing Agent (both Flowtech, Carver, MA, USA) (Figure 5 B). Finally, the aorta and inferior vena cava were ligated, and the rats were placed at 7 °C for 24 h. Constructs were explanted in toto and fixed in 3.5% formalin solution overnight and placed in PBS solution for microtomography (µCT) analysis.
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