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Development of barium-based low viscosity contrast agents for micro CT vascular casting: Application to 3D visualization of the adult mouse cerebrovasculature.

Abstract

Recent advances in three-dimensional (3D) fluorescence microscopy offer the ability to image the entire vascular network in entire organs, or even whole animals. However, these imaging modalities rely on either endogenous fluorescent reporters or involved immunohistochemistry protocols, as well as optical clearing of the tissue and refractive index matching. Conversely, X-ray-based 3D imaging modalities, such as micro CT, can image non-transparent samples, at high resolution, without requiring complicated or expensive immunolabeling and clearing protocols, or fluorescent reporters. Here, we compared two "homemade" barium-based contrast agents to the field standard, lead-containing Microfil, for micro-computed tomography (micro CT) imaging of the adult mouse cerebrovasculature. The perfusion pressure required for uniform vessel filling was significantly lower with the barium-based contrast agents compared to the polymer-based Microfil. Accordingly, the barium agents showed no evidence of vascular distension or rupture, common problems associated with Microfil. Compellingly, perfusion of an aqueous BaCl2 /gelatin mixture yielded equal or superior visualization of the cerebrovasculature by micro CT compared to Microfil. However, phosphate-containing buffers and fixatives were incompatible with BaCl2 due to the formation of unwanted precipitates. X-ray attenuation of the vessels also decreased overtime, as the BaCl2 appeared to gradually diffuse into surrounding tissues. A second, unique formulation composed of BaSO4 microparticles, generated in-house by mixing BaCl2 and MgSO4 , suffered none of these drawbacks. These microparticles, however, were unable to pass small diameter capillary vessels, conveniently labeling only the arterial cerebrovasculature. In summary, we present an affordable, robust, low pressure, non-toxic, and straightforward methodology for 3D visualization of the cerebrovasculature.

Authors+Show Affiliations

Department of Neurology, McGovern Medical School at UT Health, Houston, TX, USA.Cardiovascular Research Institute, Baylor College of Medicine, Houston, TX, USA. Department of Molecular Physiology and Biophysics, Baylor College of Medicine, Houston, TX, USA.Department of Neurology, McGovern Medical School at UT Health, Houston, TX, USA.Department of Neurology, McGovern Medical School at UT Health, Houston, TX, USA.Bruker BioSpin Corporation, Billerica, MA, USA.College of Optometry, University of Houston, Houston, TX, USA.Department of Neurology, McGovern Medical School at UT Health, Houston, TX, USA.Cardiovascular Research Institute, Baylor College of Medicine, Houston, TX, USA. Department of Molecular Physiology and Biophysics, Baylor College of Medicine, Houston, TX, USA.

Pub Type(s)

Journal Article

Language

eng

PubMed ID

31630455

Citation

Hong, Sung-Ha, et al. "Development of Barium-based Low Viscosity Contrast Agents for Micro CT Vascular Casting: Application to 3D Visualization of the Adult Mouse Cerebrovasculature." Journal of Neuroscience Research, 2019.
Hong SH, Herman AM, Stephenson JM, et al. Development of barium-based low viscosity contrast agents for micro CT vascular casting: Application to 3D visualization of the adult mouse cerebrovasculature. J Neurosci Res. 2019.
Hong, S. H., Herman, A. M., Stephenson, J. M., Wu, T., Bahadur, A. N., Burns, A. R., ... Wythe, J. D. (2019). Development of barium-based low viscosity contrast agents for micro CT vascular casting: Application to 3D visualization of the adult mouse cerebrovasculature. Journal of Neuroscience Research, doi:10.1002/jnr.24539.
Hong SH, et al. Development of Barium-based Low Viscosity Contrast Agents for Micro CT Vascular Casting: Application to 3D Visualization of the Adult Mouse Cerebrovasculature. J Neurosci Res. 2019 Oct 19; PubMed PMID: 31630455.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Development of barium-based low viscosity contrast agents for micro CT vascular casting: Application to 3D visualization of the adult mouse cerebrovasculature. AU - Hong,Sung-Ha, AU - Herman,Alexander M, AU - Stephenson,Jessica M, AU - Wu,Ting, AU - Bahadur,Ali N, AU - Burns,Alan R, AU - Marrelli,Sean P, AU - Wythe,Joshua D, Y1 - 2019/10/19/ PY - 2019/06/29/received PY - 2019/09/09/revised PY - 2019/09/23/accepted PY - 2019/10/21/entrez PY - 2019/10/21/pubmed PY - 2019/10/21/medline KW - brain KW - cerebral KW - micro CT KW - stroke KW - vasculature JF - Journal of neuroscience research JO - J. Neurosci. Res. N2 - Recent advances in three-dimensional (3D) fluorescence microscopy offer the ability to image the entire vascular network in entire organs, or even whole animals. However, these imaging modalities rely on either endogenous fluorescent reporters or involved immunohistochemistry protocols, as well as optical clearing of the tissue and refractive index matching. Conversely, X-ray-based 3D imaging modalities, such as micro CT, can image non-transparent samples, at high resolution, without requiring complicated or expensive immunolabeling and clearing protocols, or fluorescent reporters. Here, we compared two "homemade" barium-based contrast agents to the field standard, lead-containing Microfil, for micro-computed tomography (micro CT) imaging of the adult mouse cerebrovasculature. The perfusion pressure required for uniform vessel filling was significantly lower with the barium-based contrast agents compared to the polymer-based Microfil. Accordingly, the barium agents showed no evidence of vascular distension or rupture, common problems associated with Microfil. Compellingly, perfusion of an aqueous BaCl2 /gelatin mixture yielded equal or superior visualization of the cerebrovasculature by micro CT compared to Microfil. However, phosphate-containing buffers and fixatives were incompatible with BaCl2 due to the formation of unwanted precipitates. X-ray attenuation of the vessels also decreased overtime, as the BaCl2 appeared to gradually diffuse into surrounding tissues. A second, unique formulation composed of BaSO4 microparticles, generated in-house by mixing BaCl2 and MgSO4 , suffered none of these drawbacks. These microparticles, however, were unable to pass small diameter capillary vessels, conveniently labeling only the arterial cerebrovasculature. In summary, we present an affordable, robust, low pressure, non-toxic, and straightforward methodology for 3D visualization of the cerebrovasculature. SN - 1097-4547 UR - https://www.unboundmedicine.com/medline/citation/31630455/Development_of_barium-based_low_viscosity_contrast_agents_for_micro_CT_vascular_casting:_Application_to_3D_visualization_of_the_adult_mouse_cerebrovasculature L2 - https://doi.org/10.1002/jnr.24539 DB - PRIME DP - Unbound Medicine ER -