The perfusion process typically lasted for 1–2 min and was followed by perfusion with a 10% formalin solution (containing 0.01 mol/L PBS, pH 7.2–7.4, preheated to 37☌) for 3–5 min using the same perfusion parameters. Phosphate buffered saline (PBS, 0.01 mol/L, pH 7.2–7.4, preheated to 37☌) was perfused at 40–60 mmHg at a filling rate of 3–5 mL/min through the aorta until the blood outflow from the right atrium stopped. A syringe needle (Φ = 1.6 mm) was then inserted into the left ventricle, and a small slit was cut in the right atrium. We opened the chest after the mouse had been completely anesthetized. The mouse was anesthetized with an intraperitoneal injection of 5% chloral hydrate (1 mL/100 g). Based on the dataset, the thalamus was selected as an example, and its blood supply pathway was tracked and studied in this paper.Įxcept for 7 mice used in the preliminary perfusion experiments, one Kunming mouse was used in the optimized Indian-ink perfusion process. After embedding the brain with Spurr resin, a complete three-dimensional cerebrovascular dataset was acquired using Micro-Optical Sectioning Tomography (MOST). To study the entire macro and micro vascular anatomical structure network, we improved the gelatin-Indian ink vessel perfusion process based on previous work, ,, ,, and obtained a desirable perfused Kunming mouse brain. cerebrovascular connection, has not been achieved yet. However, reconstructing the connections of the blood supply pathway from the feeding vessels to the draining vessels through the capillaries in the whole brain range, i.e. Optical microscopes with three-dimensional scanning and stitching have also been used in preliminary experiments to obtain cerebrovascular dataset –, some resource have been available on the internet ( ). Vascular casts and integrated μCT, SRμ-CT and SEM have been used to image the cerebral vasculature at different resolutions. Several technologies have been developed to attempt to bridge the macro and micro vessels which form the continuous vascular network. In recent years, the study of the vascular network has become a hotspot. This leads to an urgent need for systematic research. On the other hand, microvascular imaging in a small field cannot obtain a clear view of the vessels' origins and destinations.Ĭerebral vessels play significant roles in the development and degradation of the neural network and in the process of maintaining normal brain functions. The existing techniques used to observe the macro vessels cannot visualize the fine branches of the veins, arteries and the capillary network connecting them. However, current cerebrovascular studies focus mainly either on the macro vessels of the whole brain or on the micro vessels in a small local field separately. To observe the smaller, more complex micro vessels, ex vivo two-photon laser scanning microscopy is used to image the capillary network in the cortex to a depth of 1 mm using fluorescent gelatin vessel perfusion. MRI techniques can also visualize the macro vessels in the whole mouse brain. identified and marked the major vessels in the CBA mouse using Microfil perfusion and Micro-CT imaging. The Micro-CT technique, which has been used in mice, can visualize the arterioles and venules in the whole brain. The macro vessels were identified first and were studied with the naked eye. All arteries, veins and capillaries work together to meet the demand for an uninterrupted energy supply for the brain. The topology of the cerebral vasculature, which is the energy transport corridor of the brain, can be used to study cerebral circulatory pathways –. This study provided an effective method for studying the entire macro and micro vascular networks of mouse brain simultaneously. Besides the observations of fine and complex vascular networks in the reconstructed slices and entire brain views, a representative continuous vascular tracking has been demonstrated in the deep thalamus. The voxel resolution is 0.35×0.4×2.0 µm 3 for the whole brain. With 17 days of work, an integral dataset for the entire cerebral vessels was acquired. Here, we have combined the improved gelatin-Indian ink vessel perfusion process with Micro-Optical Sectioning Tomography for imaging the vessel network of an entire mouse brain. Simultaneous vascular studies of arteries, veins and capillaries have not been achieved in the whole brain of mammals. Limited by the restrictions of the vascular markers and imaging methods, studies on cerebral vascular structure now mainly focus on either observation of the macro vessels in a whole brain or imaging of the micro vessels in a small region. The topology of the cerebral vasculature, which is the energy transport corridor of the brain, can be used to study cerebral circulatory pathways.
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