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Osmium Ammine for Staining DNA in Electron Microscopy.
Irene Masiello1, Marco Biggiogera2
1Department of Biology and Biotechnology, Laboratory of Cell Biology and Neurobiology, University of Pavia, Via Ferrata 9, 27100, Pavia, Italy.
Methods in Molecular Biology (Clifton, N.J.)
|February 4, 2017
Summary
Osmium ammine staining precisely detects DNA in cell nuclei using a Feulgen-type reaction. This method, suitable for electron microscopy, involves acid hydrolysis and Schiff-type reagent binding for ultrathin sections.
Area of Science:
- Cell Biology
- Microscopy Techniques
- Biochemistry
Background:
- Osmium ammine staining is a widely used technique in electron microscopy cytochemistry.
- It specifically detects DNA within the cell nucleus.
Purpose of the Study:
- To detail the osmium ammine staining procedure for ultrathin sections.
- To describe variations in reagent preparation for different section types (acrylic and epoxy).
Main Methods:
- The technique is a Feulgen-type reaction.
- It involves acid hydrolysis to generate aldehyde groups on DNA.
- These aldehydes bind to osmium ammine, a Schiff-type reagent.
Main Results:
- Osmium ammine provides specific detection of DNA in the cell nucleus.
- The staining procedure is optimized for ultrathin sections.
- Modified reagent preparation protocols are provided for acrylic and epoxy sections.
Conclusions:
- Osmium ammine staining is a reliable method for DNA detection in EM cytochemistry.
- The described procedures enhance its applicability to various embedding media.
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