Related Experiment Video
Updated: Aug 27, 2025
Preparation and Immunostaining of Myelinating Organotypic Cerebellar Slice Cultures
Published on: March 20, 2019
Staining Methods for Normal and Regenerative Myelin in the Nervous System
Óscar D García-García1,2, Tamara Weiss3, Jesús Chato-Astrain1,2
1Department of Histology (Tissue Engineering Group), Faculty of Medicine, University of Granada, Granada, Spain.
Insights
This chapter details four histological methods for myelin evaluation in peripheral nerve tissue. Techniques include Luxol fast blue (LFB) with MCOLL, osmium tetroxide, immunofluorescence, and FluoroMyelin™ for comprehensive myelin identification.
Area of Science:
- Neuroscience
- Histology
- Biochemistry
Background:
- Myelin identification is crucial for understanding peripheral nerve structure and function.
- Histochemical and fluorescence microscopy are key tools for myelin visualization.
Purpose of the Study:
- To describe four distinct histological methods for evaluating myelin in peripheral nerve tissue sections.
- To provide researchers with versatile techniques for myelin analysis in various experimental contexts.
Main Methods:
- Luxol fast blue (LFB) combined with modified Picrosirius staining (MCOLL) for myelin, collagen, and nuclei.
- Osmium tetroxide staining for myelin and other lipids.
- Immunofluorescence detection of myelin proteins.
- FluoroMyelin™ stains for rapid myelin detection in cryosections.
Main Results:
- The MCOLL method offers an integrated view of histology, collagen, and myelin in embedded or cryosectioned samples.
- Osmium tetroxide provides a permanent stain for myelin and lipids.
- Immunofluorescence allows co-detection of myelin proteins with other targets.
- FluoroMyelin™ enables quick myelin assessment within immunofluorescence panels.
Conclusions:
- A range of histological and fluorescence-based methods are available for accurate myelin identification.
- These techniques support diverse experimental approaches in peripheral nerve research.
- The described methods facilitate detailed analysis of myelin content and associated structures.
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.
Related Concept Videos
01:25Nervous Tissue: Myelin
Schwann cells begin to form myelin sheaths around axons during fetal development. They wrap around a small...
01:15Neurogenesis and Regeneration of Nervous Tissue