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Detection of elastin by immunoelectronmicroscopy. A comparison of different procedures
D Daga-Gordini1, G M Bressan, I Castellani
1Institute of Histology and Embryology, University of Padova, Italy.
Histochemistry
|January 1, 1987
Summary
Immunoelectronmicroscopy revealed elastin components in chick aorta. Artifacts in the peroxidase-antiperoxidase method caused false positives in microfibrils and cell surfaces, unlike immunogold labeling.
Area of Science:
- Cell Biology
- Biochemistry
- Immunohistochemistry
Background:
- Elastin is a crucial protein in the aorta, providing elasticity.
- Understanding elastin's precise localization is vital for cardiovascular research.
- Immunoelectronmicroscopy is a key technique for visualizing cellular components.
Purpose of the Study:
- To identify and map elastin components within the chick aorta.
- To evaluate different immunoelectronmicroscopic techniques for elastin localization.
- To differentiate true elastin localization from potential artifacts.
Main Methods:
- Utilized three immunoelectronmicroscopic methods: peroxidase-antiperoxidase (PAP), immunoferritin, and immunogold.
- Employed affinity-purified antibodies against chick tropoelastin.
- Performed pre-embedding and on-section labeling on Epon and Lowicryl embedded tissues.
Main Results:
- The PAP method intensely stained amorphous elastin and microfibrils, with slight cell surface staining.
- Immunogold labeling exclusively and strongly decorated amorphous elastin, leaving microfibrils unlabeled.
- Artifacts due to diffusion of peroxidase reaction products explained the PAP method's microfibril and cell surface staining.
Conclusions:
- Immunogold labeling is a more reliable method for accurate fine mapping of elastin components.
- The PAP method, under the conditions used, produced artifacts that mislocalized elastin components.
- Careful selection of immunological reagents and procedures is essential for valid ultrastructural studies of elastin.