Translational Stroke Research

, Volume 3, Supplement 1, pp 174–179

Micro-Computed Tomography for Hemorrhage Disruption of Mouse Brain Vasculature

  • Bohua Xie
  • Peng Miao
  • Yuhao Sun
  • Yongting Wang
  • Guo-Yuan Yang
Original Article

Abstract

The use of genetic engineering to develop important neuropathological mouse models has made cerebrovascular imaging essential for the investigation of numerous brain disorders, especially cerebrovascular disorders, such as aneurysms, arteriovenous malformations, and ischemic and hemorrhagic stroke. New laboratory-based X-ray microimagers exist that provide easy access, reliable operation, and performance previously found only in synchrotron-based instruments. Here, we reported a novel approach using such a system to detect intracerebral hemorrhage and resultant cerebrovascular pathology. Adult male C57BL/6 mice (n = 12) underwent 30 μl autologous blood injection into the right basal ganglia region. After sacrificing the animals and vascular perfusion with Microfil® MV-122 Yellow to opacify vascular and microvascular structures, the brain was post-fixed and partially hydrated for 3D imaging with a MicroXCT-400® at 30 KeV and 2-μm resolution. Tomographic reconstruction of high-resolution microimages was accomplished with Amira® software. High-quality 3D images included cerebrocortical microvessels, the circle of Willis, the sagittal sinus, transverse sinus, and other arterial and venous systems. In the ipsilateral hemisphere, there clearly were early-stage vasodilatation and later-stage neovascularization. Very high-resolution, laboratory-based, X-ray micro-CT contrast imaging can accomplish sensitive quantifications of normal and pathological small cerebrovascular changes, especially in hemorrhagic stroke and subsequent hemorrhage-induced neovascularization.

Keywords

Brain Hemorrhage Imaging Micro-XCT Mouse Vasculature 

Abbreviations

ACA

Anterior cerebral artery

BA

Basal artery

ICA

Internal carotid artery

ICH

Intracerebral hemorrhage

MCA

Middle cerebral artery

Micro-XCT

Micro-X-ray computerized tomography

3D

Three dimensional

VOI

Volume of interest

References

  1. 1.
    Elliott J, Smith M. The acute management of intracerebral hemorrhage: a clinical review. Anesth Analg. 2010;110(5):1419–27. Epub 2010/03/25.PubMedCrossRefGoogle Scholar
  2. 2.
    Woo D, Broderick JP. Spontaneous intracerebral hemorrhage: epidemiology and clinical presentation. Neurosurg Clin N Am. 2002;13(3):265–79. v. Epub 2002/12/19.PubMedCrossRefGoogle Scholar
  3. 3.
    Mendelow AD, Gregson BA, Fernandes HM, Murray GD, Teasdale GM, Hope DT, et al. Early surgery versus initial conservative treatment in patients with spontaneous supratentorial intracerebral haematomas in the International Surgical Trial in Intracerebral Haemorrhage (STICH): a randomised trial. Lancet. 2005;365(9457):387–97. Epub 2005/02/01.PubMedGoogle Scholar
  4. 4.
    Mayer SA. Recombinant activated factor VII for acute intracerebral hemorrhage. Stroke. 2007;38(2 Suppl):763–7. Epub 2007/01/31.PubMedCrossRefGoogle Scholar
  5. 5.
    Mayer SA, Brun NC, Begtrup K, Broderick J, Davis S, Diringer MN, et al. Recombinant activated factor VII for acute intracerebral hemorrhage. N Engl J Med. 2005;352(8):777–85. Epub 2005/02/25.PubMedCrossRefGoogle Scholar
  6. 6.
    Xi G, Keep RF, Hoff JT. Mechanisms of brain injury after intracerebral haemorrhage. Lancet Neurol. 2006;5(1):53–63. Epub 2005/12/20.PubMedCrossRefGoogle Scholar
  7. 7.
    Wagner KR, Sharp FR, Ardizzone TD, Lu A, Clark JF. Heme and iron metabolism: role in cerebral hemorrhage. J Cereb Blood Flow Metab. 2003;23(6):629–52. Epub 2003/06/11.PubMedCrossRefGoogle Scholar
  8. 8.
    Xue M, Mikliaeva EI, Casha S, Zygun D, Demchuk A, Yong VW. Improving outcomes of neuroprotection by minocycline: guides from cell culture and intracerebral hemorrhage in mice. Am J Pathol. 2010;176(3):1193–202. Epub 2010/01/30.PubMedCrossRefGoogle Scholar
  9. 9.
    Hwang BY, Appelboom G, Ayer A, Kellner CP, Kotchetkov IS, Gigante PR, et al. Advances in neuroprotective strategies: potential therapies for intracerebral hemorrhage. Cerebrovasc Dis. 2011;31(3):211–22. Epub 2010/12/24.PubMedCrossRefGoogle Scholar
  10. 10.
    Zazulia AR, Diringer MN, Derdeyn CP, Powers WJ. Progression of mass effect after intracerebral hemorrhage. Stroke. 1999;30(6):1167–73. Epub 1999/06/04.PubMedCrossRefGoogle Scholar
  11. 11.
    Walker E, Shen F, Young W, Su H. Cerebrovascular casting of the adult mouse for 3D imaging and morphological analysis. J Vis Exp. 2011;(57):e2958.Google Scholar
  12. 12.
    Plouraboué F, Cloetens P, Fonta C, Steyer A, Lauwers F, MARC-VERGNES JP. X-ray high-resolution vascular network imaging. J Microsc. 2004;215(2):139–48.PubMedCrossRefGoogle Scholar
  13. 13.
    Liao PS, Chen TS, Chung PC. A fast algorithm for multilevel thresholding. J Inf Sci Eng. 2001;17(5):713–28.Google Scholar
  14. 14.
    Pathak AP, Kim E, Zhang J, Jones MV. Three-dimensional imaging of the mouse neurovasculature with magnetic resonance microscopy. PLoS One. 2011;6(7):e22643.PubMedCrossRefGoogle Scholar
  15. 15.
    de Crespigny A, Bou-Reslan H, Nishimura MC, Phillips H, Carano RAD, D'Arceuil HE. 3D micro-CT imaging of the postmortem brain. J Neurosci Methods. 2008;171(2):207–13.PubMedCrossRefGoogle Scholar
  16. 16.
    Beckmann F, Heise K, Kölsch B, Bonse U, Rajewsky M, Bartscher M, et al. Three-dimensional imaging of nerve tissue by x-ray phase-contrast microtomography. Biophys J. 1999;76(1):98–102.PubMedCrossRefGoogle Scholar
  17. 17.
    Risser L, Plouraboué F, Cloetens P, Fonta C. A 3D-investigation shows that angiogenesis in primate cerebral cortex mainly occurs at capillary level. Int J Dev Neurosci. 2009;27(2):185–96.PubMedCrossRefGoogle Scholar
  18. 18.
    Metscher B. MicroCT for comparative morphology: simple staining methods allow high-contrast 3D imaging of diverse non-mineralized animal tissues. BMC Physiol. 2009;9(1):11.PubMedCrossRefGoogle Scholar
  19. 19.
    Metscher BD. MicroCT for developmental biology: a versatile tool for high-contrast 3D imaging at histological resolutions. Dev Dyn. 2009;238(3):632–40.PubMedCrossRefGoogle Scholar
  20. 20.
    Kim JS, Min J, Recknagel AK, Riccio M, Butcher JT. Quantitative three-dimensional analysis of embryonic chick morphogenesis via microcomputed tomography. The Anatomical Record: Advances in Integrative Anatomy and Evolutionary Biology. 2010;294:1–10.CrossRefGoogle Scholar
  21. 21.
    Holdsworth DW, Thornton MM. Micro-CT in small animal and specimen imaging. Trends Biotechnol. 2002;20(8):S34–S9.CrossRefGoogle Scholar
  22. 22.
    Simopoulos DN, Gibbons SJ, Malysz J, Szurszewski JH, Farrugia G, Ritman EL, et al. Corporeal structural and vascular micro architecture with X-ray micro computerized tomography in normal and diabetic rabbits: histopathological correlation. J Urol. 2001;165(5):1776–82.PubMedCrossRefGoogle Scholar
  23. 23.
    Wilson SH, Herrmann J, Lerman LO, Holmes DR, Napoli C, Ritman EL, et al. Simvastatin preserves the structure of coronary adventitial vasa vasorum in experimental hypercholesterolemia independent of lipid lowering. Circulation. 2002;105(4):415–8.PubMedCrossRefGoogle Scholar
  24. 24.
    Yang GY, Betz AL, Chenevert TL, Brunberg JA, Hoff JT. Experimental intracerebral hemorrhage: relationship between brain edema, blood flow, and blood–brain barrier permeability in rats. J Neurosurg. 1994;81(1):93–102.PubMedCrossRefGoogle Scholar
  25. 25.
    Lee TC, Kashyap RL, Chu CN. Building skeleton models via 3-D medial surface/axis thinning algorithms. CVGIP: Graphical Model and Image Processing. 1994;56(6):462–78.CrossRefGoogle Scholar
  26. 26.
    Marxen M, Thornton MM, Chiarot CB, Klement G, Koprivnikar J, Sled JG, et al. MicroCT scanner performance and considerations for vascular specimen imaging. Medical Ph1ysics. 2004;31:305.CrossRefGoogle Scholar
  27. 27.
    Bolland BJ, Kanczler JM, Dunlop DG, Oreffo RO. Development of in vivo muCT evaluation of neovascularisation in tissue engineered bone constructs. Bone. 2008;43(1):195–202. Epub 2008/04/22.PubMedCrossRefGoogle Scholar
  28. 28.
    Chu M, Hu X, Lu S, Gan Y, Li P, Guo Y, et al. Focal cerebral ischemia activates neurovascular restorative dynamics in mouse brain. Front Biosci (Elite Ed). 2012;4:1926.Google Scholar

Copyright information

© Springer Science+Business Media, LLC 2012

Authors and Affiliations

  • Bohua Xie
    • 1
    • 2
  • Peng Miao
    • 1
    • 5
  • Yuhao Sun
    • 1
    • 4
  • Yongting Wang
    • 1
  • Guo-Yuan Yang
    • 1
    • 3
    • 6
  1. 1.School of Biomedical Engineering and Med-X Research InstituteShanghai Jiao Tong UniversityShanghaiChina
  2. 2.School of Life Science and BiotechnologyShanghai Jiao Tong UniversityShanghaiChina
  3. 3.Department of Neurology, Ruijin Hospital, School of MedicineShanghai Jiao Tong UniversityShanghaiChina
  4. 4.Department of Neurosurgery, Ruijin Hospital, School of MedicineShanghai Jiao Tong UniversityShanghaiChina
  5. 5.School of Communication and Information EngineeringShanghai UniversityShanghaiChina
  6. 6.Neuroscience and Neuroengineering Center, Med-X Research Institute and School of Biomedical EngineeringShanghai Jiao Tong UniversityShanghaiChina

Personalised recommendations