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Diaminobenzidine as a myelin stain in semithin plastic sections
S K Krueger1, D E Phillips, M M Frederick
1Department of Biology, Montana State University, Bozeman 59717-0346, USA.
Abstract:
A diaminobenzidine (DAB) stain for myelin in glutaraldehyde fixed, osmicated, semithin epoxy sections is described. One or 1.5 microm sections, dried onto slides, are first etched with a 1:2 dilution of saturated sodium ethoxide:absolute ethanol, then incubated in 0.05% aqueous DAB with 0.01% hydrogen peroxide. DAB specifically stains osmium fixed myelinated nerve fibers. This permits high resolution light microscopic study of myelinated nerve fibers in semithin sections of tissues that also can be studied by electron microscopy.
Insights
A new diaminobenzidine (DAB) stain effectively highlights myelin in semithin epoxy sections. This method enables high-resolution light microscopy of myelinated nerve fibers for detailed tissue analysis.
Area of Science:
- Neuroscience
- Histology
- Microscopy
Background:
- Myelin staining is crucial for studying myelinated nerve fibers.
- Existing methods may not be compatible with subsequent electron microscopy.
- Semithin epoxy sections offer potential for correlative light and electron microscopy.
Purpose of the Study:
- To develop a reliable diaminobenzidine (DAB) staining protocol for myelin in semithin epoxy sections.
- To enable high-resolution light microscopic visualization of myelinated nerve fibers in tissues prepared for electron microscopy.
Main Methods:
- Utilized glutaraldehyde fixation and osmium tetroxide postfixation.
- Employed semithin (1-1.5 micrometer) epoxy sections.
- Developed a specific staining procedure involving etching with sodium ethoxide/ethanol and incubation with DAB and hydrogen peroxide.
Main Results:
- Diaminobenzidine (DAB) specifically stained osmium-fixed myelinated nerve fibers.
- The stain provided clear visualization of myelin sheaths in semithin sections.
- The stained sections remained suitable for subsequent electron microscopy.
Conclusions:
- The described DAB staining method is effective for myelin visualization in semithin epoxy sections.
- This technique facilitates correlative light and electron microscopy studies of myelinated nerve fibers.
- It offers a valuable tool for high-resolution light microscopic analysis in neuroscience research.
