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Immunohistochemical analysis of various serum proteins in living mouse thymus with “in vivo cryotechnique”

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Abstract

It has been difficult to clarify the precise localizations of soluble serum proteins in thymic tissues of living animals with conventional immersion- or perfusion-fixation followed by alcohol dehydration owing to ischemia and anoxia. In this study, “in vivo cryotechnique” (IVCT) followed by freeze-substitution fixation was performed to examine the thymic structures of living mice and immunolocalizations of intrinsic or extrinsic serum proteins, which were albumin, immunoglobulin G1 (IgG1), IgA, and IgM, as well as intravenously injected bovine serum albumin (BSA). Mouse albumin was more clearly immunolocalized in blood vessels and interstitial matrices of the thymic cortex than in tissues prepared by the conventional methods. The immunoreactivities of albumin and IgG1 were stronger than those of IgA and IgM in the interstitium of subcapsular cortex. The injected BSA was time-dependently immunolocalized in blood vessels and the interstitium of corticomedullary areas at 3.5 h after its injection, and then gradually diffused into the interstitium of the whole cortex at 6 h and 12 h. Thus, IVCT revealed definite immunolocalizations of serum albumin and IgG1 in the interstitium of thymus of living mice, indicating different accessibility of serum proteins from the corticomedullary areas, not from the subcapsular cortex of living animals, depending on various molecular sizes and concentrations.

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Authors and Affiliations

  1. Department of Anatomy and Molecular Histology, Interdisciplinary Graduate School of Medicine and Engineering, University of Yamanashi, 1110 Shimokato, Chuo, Yamanashi, 409-3898, Japan

    Yuqin Bai, Bao Wu, Nobuo Terada, Yurika Saitoh, Nobuhiko Ohno, Sei Saitoh & Shinichi Ohno

Authors
  1. Yuqin Bai
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  2. Bao Wu
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  3. Nobuo Terada
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  4. Yurika Saitoh
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  5. Nobuhiko Ohno
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  6. Sei Saitoh
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  7. Shinichi Ohno
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Corresponding author

Correspondence to Shinichi Ohno.

Additional information

Dr. Y. Bai was a research fellow from the Department of Pathology, Faculty of Medicine, Chifeng University, Inner Mongolia 024000, China, while this work was in progress at the University of Yamanashi.

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Bai, Y., Wu, B., Terada, N. et al. Immunohistochemical analysis of various serum proteins in living mouse thymus with “in vivo cryotechnique”. Med Mol Morphol 45, 129–139 (2012). https://doi.org/10.1007/s00795-011-0549-0

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  • DOI: https://doi.org/10.1007/s00795-011-0549-0

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