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Electron microscopy : use of lectin-gold after embedding
1Department of Histology, Anatomy Centre, Kreuzbergring 36, Göttingen, Germany.
Methods in Molecular Medicine
|March 5, 2011
Summary
Choosing the right tissue fixative is crucial for successful postembedding gold-lectin histochemistry. A compromise fixative, like formaldehyde and glutaraldehyde, is recommended over osmium tetroxide to preserve tissue structure and lectin binding sites.
Area of Science:
- Histochemistry
- Biochemistry
- Microscopy
Background:
- Postembedding gold-lectin histochemistry requires specific tissue fixation methods.
- Standard fixation protocols may not preserve ultrastructure or lectin binding sites effectively.
- Glycoconjugates must be strongly bound to prevent washout during tissue processing.
Purpose of the Study:
- To evaluate the properties of ideal fixatives for postembedding gold-lectin histochemistry.
- To identify suitable fixative compromises for preserving tissue ultrastructure and lectin binding.
- To determine the impact of different fixatives on lectin binding site reactivity.
Main Methods:
- Review of fixation requirements for postembedding gold-lectin histochemistry.
- Analysis of common fixatives, including formaldehyde, glutaraldehyde, and osmium tetroxide.
- Assessment of fixative effects on tissue ultrastructure and lectin binding site integrity.
Main Results:
- No single fixative perfectly meets all requirements for postembedding gold-lectin histochemistry.
- A mixture of 4% formaldehyde and 0.5% glutaraldehyde in phosphate buffer (pH 7.35) is a commonly used compromise.
- Osmium tetroxide is generally unsuitable as it can interfere with lectin binding site reactivity.
Conclusions:
- Fixative selection for postembedding gold-lectin histochemistry involves balancing competing requirements.
- Formaldehyde-glutaraldehyde mixtures offer a viable compromise for preserving tissue and lectin binding.
- Avoiding osmium tetroxide is recommended to ensure optimal lectin histochemistry results.
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