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Gold toning preserves integrity of silver enhanced immunogold particles during osmium tetroxide treatment for demonstration of a biogenic amine

Lookup NU author(s): Dr Gerd Leitinger

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Abstract

We describe a protocol that enhances immunolabelling of nervous tissue for ultrastructural study. Insect tissue is fixed, sectioned, and labelled with a polyclonal antiserum against serotonin and a secondary antibody conjugated with 1 nm colloidal gold. The gold particles are silver-enhanced to ease detection and then protected by gold toning. Finally, the tissue is post fixed in glutaraldehyde fixative followed by osmium tetroxide and further processed for electron microscopy. We demonstrated on insect nervous tissue that gold toning protects marker particles from the influence of osmium tetroxide, Use of buffered solutions throughout the protocol led to well preserved ultrastructural details, and marker particle size was nor reduced with a short gold toning time. We also suggest use of this protocol for vertebrate or other invertebrate tissue. (C) 2000 Elsevier Science B.V. All rights reserved.

Publication metadata

Author(s): Leitinger G, Pabst MA, Kral K

Publication type: Article

Publication status: Published

Journal: Brain Research Protocols

Year: 2000

Volume: 5

Issue: 1

Pages: 30-38

ISSN (print): 1385-299X

ISSN (electronic): 1872-809X

Publisher: Elsevier

URL: http://dx.doi.org/10.1016/S1385-299X(99)00049-5

DOI: 10.1016/S1385-299X(99)00049-5

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