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Staining Methods for Normal and Regenerative Myelin in the Nervous System

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Histochemistry of Single Molecules

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2566))

Abstract

Histochemical and fluorescence-based techniques enable the specific identification of myelin by bright-field or fluorescence microscopy. In this chapter, we describe four histological methods for the evaluation of myelin on peripheral nerve tissue sections. The first method combines the Luxol fast blue (LFB) technique with a modified Picrosirius staining contrasted with Harris hematoxylin, called MCOLL. This method simultaneously stains myelin, collagen fibers, and cell nuclei, thus giving an integrated overview of the histology, collagen network, and myelin content of the tissue in paraffin-embedded or cryosectioned samples. Secondly, we describe the osmium tetroxide method, which provides a permanent positive reaction for myelin as well as other lipids present in the tissue. The third method is the immunofluorescence-based detection of myelin proteins that allows to combine information about their expression status with other proteins of interest. Finally, the FluoroMyelin™ stains enable a fast detection of the myelin content that can be easily implemented in immunofluorescence staining panels for cryosectioned tissues. Together, this chapter provides a variety of methods to accurately identify myelin in different experimental approaches.

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Acknowledgments

Methods described in this chapter were supported by the Spanish Plan Nacional de Investigación Científica, Desarrollo e Innovación Tecnológica, Ministerio de Economía y Competitividad (Instituto de Salud Carlos III), Grants No FIS PI17-0393, FIS PI20-0318 co-financed by the “Fondo Europeo de Desarrollo Regional ERDF-FEDER European Union”; Grant No P18-RT-5059 by Plan Andaluz de Investigación, Desarrollo e Innovación (PAIDI 2020), Consejería de Transformación Económica, Industria, Conocimiento y Universidades, Junta de Andalucía, España; and Grant A-CTS-498-UGR18 by Programa Operativo FEDER Andalucía 2014–2020, Universidad de Granada, Junta de Andalucía, España, co-funded by ERDF-FEDER, the European Union. Authors are grateful to Ms. Fabiola Bermejo Casares for her technical support and Prof. Ariane Ruyffelaert for proofreading.

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Author notes

  1. Óscar D. García-García and Tamara Weiss contributed equally to this chapter.

Authors and Affiliations

  1. Department of Histology (Tissue Engineering Group), Faculty of Medicine, University of Granada, Granada, Spain

    Óscar D. García-García, Jesús Chato-Astrain & Víctor Carriel

  2. Instituto de Investigación Biosanitaria, Ibs.GRANADA, Granada, Spain

    Óscar D. García-García, Jesús Chato-Astrain & Víctor Carriel

  3. Department of Plastic, Reconstructive and Aesthetic Surgery, Medical University of Vienna, Vienna, Austria

    Tamara Weiss

  4. Dipartimento di Scienze Cliniche e Biologiche, Università di Torino, Torino, Italy

    Stefania Raimondo

  5. Neuroscience Institute Cavalieri Ottolenghi (NICO), Torino, Italy

    Stefania Raimondo

Authors
  1. Óscar D. García-García
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  2. Tamara Weiss
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  3. Jesús Chato-Astrain
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  4. Stefania Raimondo
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  5. Víctor Carriel
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Corresponding author

Correspondence to Víctor Carriel .

Editor information

Editors and Affiliations

  1. Department of Biology and Biotechnology, University of Pavia, PAVIA, Italy

    Carlo Pellicciari

  2. Department of Biology and Biotechnology, University of Pavia, PAVIA, Italy

    Marco Biggiogera

  3. Department of Neurosciences, Biomedicine and Movement Sciences, University of Verona, VERONA, Italy

    Manuela Malatesta

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© 2023 The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature

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García-García, Ó.D., Weiss, T., Chato-Astrain, J., Raimondo, S., Carriel, V. (2023). Staining Methods for Normal and Regenerative Myelin in the Nervous System. In: Pellicciari, C., Biggiogera, M., Malatesta, M. (eds) Histochemistry of Single Molecules. Methods in Molecular Biology, vol 2566. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-2675-7_15

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  • DOI: https://doi.org/10.1007/978-1-0716-2675-7_15

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-2674-0

  • Online ISBN: 978-1-0716-2675-7

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