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Pimonidazole Adduct Immunohistochemistry in the Rat Kidney: Detection of Tissue Hypoxia

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Kidney Research

Part of the book series: Methods in Molecular Biology ((MIMB,volume 466))

Abstract

Immunohistochemistry for pimonidazole adducts serves to define hypoxia within tissues. For this purpose, pimonidazole is delivered in vivo, binds to thiol groups at oxygen tensions below 10 mmHg, and is visualized with help of commercially available anti-pimonidazole antibodies. Renal parenchymal oxygen distribution is highly variable under normal conditions and during acute renal failure and chronic renal disorders. Pimonidazole immunostaining clearly helps in delineating hypoxic regions within the kidneys, but technical pitfalls should be taken into account. In particular, tissue fixation by in vivo perfusion is strongly recommended in order to eliminate artificial staining, because immersion fixation per se can promote a hypoxic environment within kidney tissue.

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Acknowledgment

We thank Professor Sebastian Bachmann for advice in renal perfusion fixation and for reviewing the manuscript.

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Authors and Affiliations

  1. CharitéUniversitaetsmedizin, Nephrology and Medical Intensive Care, Berlin, Germany

    Christian Rosenberger

  2. Beth Israel Deaconess Medical Centre and Harvard Medical School, Pathology, Boston, MA, USA

    Seymour Rosen

  3. Department of Anatomy, Humboldt University, Berlin, Germany

    Alexander Paliege

  4. Hadassah University Hospital Mt. Scopus and The Hebrew Medical School, Medicine, Jerusalem, Israel

    Samuel N. Heyman

Authors
  1. Christian Rosenberger
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  2. Seymour Rosen
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  3. Alexander Paliege
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  4. Samuel N. Heyman
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    © 2009 Humana Press, a part of Springer Science+Business Media, LLC

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    Rosenberger, C., Rosen, S., Paliege, A., Heyman, S.N. (2009). Pimonidazole Adduct Immunohistochemistry in the Rat Kidney: Detection of Tissue Hypoxia. In: Becker, G., Hewitson, T. (eds) Kidney Research. Methods in Molecular Biology, vol 466. Humana Press. https://doi.org/10.1007/978-1-59745-352-3_12

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    • DOI: https://doi.org/10.1007/978-1-59745-352-3_12

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    • Publisher Name: Humana Press

    • Print ISBN: 978-1-58829-945-1

    • Online ISBN: 978-1-59745-352-3

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