Abstract
In this chapter, we present the application of “in vivo cryotechnique” (IVCT) to examination of time-dependent topographical changes of leaking proteins from blood vessels in the mouse cerebellum, to assess the blood–brain barrier (BBB). The distribution of leaking serum proteins was compared by various cryotechniques. The cryofixed cerebellar tissues were processed for the freeze-substitution method and paraffin embedding. Serial deparaffinized sections were immunostained by anti-mouse immunoglobulin G (IgG) or albumin antibody. By combination of IVCT, serum IgG and albumin were clearly localized inside of cerebellar blood vessels. In anoxic cerebellar tissues, which were partially removed from brains in the mouse skull and quickly frozen in the isopentane–propane within a minute, serum IgG and albumin were diffusely immunostained around the blood capillaries, showing leakage of the serum components through the BBB changes. Thus, IVCT revealed in vivo localization of serum components in mouse brains.
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Terada, N., Saitoh, Y., Ohno, N., Zea-Aragón, Z., Ohno, S. (2016). Application of “In Vivo Cryotechnique” to Immunohistochemical Analyses for Effects of Anoxia on Serum Immunoglobulin and Albumin Leakage Through Blood–Brain Barrier in Mouse Cerebellum. In: Ohno, S., Ohno, N., Terada, N. (eds) In Vivo Cryotechnique in Biomedical Research and Application for Bioimaging of Living Animal Organs. Springer, Tokyo. https://doi.org/10.1007/978-4-431-55723-4_32
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DOI: https://doi.org/10.1007/978-4-431-55723-4_32
Publisher Name: Springer, Tokyo
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