Dataset Information

Vascular Casting of Adult and Early Postnatal Mouse Lungs for Micro-CT Imaging.

ABSTRACT: Blood vessels form intricate networks in 3-dimensional space. Consequently, it is difficult to visually appreciate how vascular networks interact and behave by observing the surface of a tissue. This method provides a means to visualize the complex 3-dimensional vascular architecture of the lung. To accomplish this, a catheter is inserted into the pulmonary artery and the vasculature is simultaneously flushed of blood and chemically dilated to limit resistance. Lungs are then inflated through the trachea at a standard pressure and the polymer compound is infused into the vascular bed at a standard flow rate. Once the entire arterial network is filled and allowed to cure, the lung vasculature may be visualized directly or imaged on a micro-CT (µCT) scanner. When performed successfully, one can appreciate the pulmonary arterial network in mice ranging from early postnatal ages to adults. Additionally, while demonstrated in the pulmonary arterial bed, this method can be applied to any vascular bed with optimized catheter placement and endpoints.

SUBMITTER: Knutsen RH

PROVIDER: S-EPMC8284398 | biostudies-literature | 2020 Jun

REPOSITORIES: biostudies-literature

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Vascular Casting of Adult and Early Postnatal Mouse Lungs for Micro-CT Imaging.

Knutsen Russell H RH Gober Leah M LM Sukinik Joseph R JR Donahue Danielle R DR Kronquist Elise K EK Levin Mark D MD McLean Sean E SE Kozel Beth A BA

Journal of visualized experiments : JoVE 20200620 160

Blood vessels form intricate networks in 3-dimensional space. Consequently, it is difficult to visually appreciate how vascular networks interact and behave by observing the surface of a tissue. This method provides a means to visualize the complex 3-dimensional vascular architecture of the lung. To accomplish this, a catheter is inserted into the pulmonary artery and the vasculature is simultaneously flushed of blood and chemically dilated to limit resistance. Lungs are then inflated through th ...[more]

PMID: 32628170

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Project description:BackgroundMicrovascular visualization is crucial in understanding the mechanisms of several pathologies. For instance, visualization of the tumor microenvironment is important in understanding angiogenesis and role in cancer progression. Visualization would provide insights to cancer diagnosis, predicting metastatic growth, and evaluating therapeutic protocols. Similarly, understanding the microvascular network could be beneficial for study of degenerative diseases and tissue repair. The use of microscale computed tomography (micro-CT) and vascular casting agents provides high-resolution images of tissue vasculature in volumetric space. The purpose of this research was to compare a selection of commercially available contrast agents to determine the optimal solution for vascular visualization.MethodsA population of 16 female nude athymic mice (Charles Rivers Laboratories) were implanted with MDA-MB-231 breast cancer cells (ATCC) orthotopically in the lower left mammary fat pad to investigate the tumor microenvironment. Once tumors reach sufficient size, animals were equally divided into four groups based on the micro-CT agent to be administered, namely, control (no contrast agent), barium sulfate (BaSO4), Vascupaint, or Microfil. Animals were anesthetized prior to transcarotid micro-cannulation to infuse 2 mL of the specific contrast agent for intravascular distribution throughout the animal. The jugular vein on the other side of the carotid artery was opened to drain blood flow. Following successful perfusion, animals and extracted organs underwent high-resolution micro-CT scanning (OI/CT, MILabs). Images were reconstructed and analyzed using analysis software to extract mean intensity signals.ResultsPreliminary post-mortem micro-CT results reveal Vascupaint and BaSO4 are useful for microvascular visualization. Both Vascupaint and BaSO4 produced significant contrast-enhanced micro-CT image enhancement in the brain (3.39±0.93 and 6.27±3.78, respectively) and kidney (12.85±1.98 and 32.87±10.03, respectively) as compared to Microfil (0.22±0.07 and 0.91±0.63, respectively; P<0.05). For the various contrast agents, there were no differences in image enhancement from the liver, spleen, or tumor tissue (P>0.21). Moreover, use of Vascupaint and BaSO4 allowed for visualization of smaller microvascular structures with average diameters of 20.54±4.15 and 25.82±3.75 µm, which were smaller compared to the 91.66±24.91 µm measurements from Microfil-enhanced micro-CT images (P<0.004).ConclusionsOur study suggests that the use of Vascupaint and BaSO4 is more than sufficient for ex vivo visualization of microvascular structures with contrast-enhanced micro-CT imaging as these contrast agents more effectively perfused smaller blood vessels.

Project description:Micro-CT is widely used for small animal imaging in preclinical studies of cardiopulmonary disease, but further development is needed to improve spatial resolution, temporal resolution, and material contrast. We present a technique for visualizing the changing distribution of iodine in the cardiac cycle with dual source micro-CT.The approach entails a retrospectively gated dual energy scan with optimized filters and voltages, and a series of computational operations to reconstruct the data. Projection interpolation and five-dimensional bilateral filtration (three spatial dimensions + time + energy) are used to reduce noise and artifacts associated with retrospective gating. We reconstruct separate volumes corresponding to different cardiac phases and apply a linear transformation to decompose these volumes into components representing concentrations of water and iodine. Since the resulting material images are still compromised by noise, we improve their quality in an iterative process that minimizes the discrepancy between the original acquired projections and the projections predicted by the reconstructed volumes. The values in the voxels of each of the reconstructed volumes represent the coefficients of linear combinations of basis functions over time and energy. We have implemented the reconstruction algorithm on a graphics processing unit (GPU) with CUDA. We tested the utility of the technique in simulations and applied the technique in an in vivo scan of a C57BL∕6 mouse injected with blood pool contrast agent at a dose of 0.01 ml∕g body weight. Postreconstruction, at each cardiac phase in the iodine images, we segmented the left ventricle and computed its volume. Using the maximum and minimum volumes in the left ventricle, we calculated the stroke volume, the ejection fraction, and the cardiac output.Our proposed method produces five-dimensional volumetric images that distinguish different materials at different points in time, and can be used to segment regions containing iodinated blood and compute measures of cardiac function.We believe this combined spectral and temporal imaging technique will be useful for future studies of cardiopulmonary disease in small animals.

Dataset Information

Vascular Casting of Adult and Early Postnatal Mouse Lungs for Micro-CT Imaging.

Publications

Vascular Casting of Adult and Early Postnatal Mouse Lungs for Micro-CT Imaging.

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