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Journal of Histochemistry and Cytochemistry, Vol. 48, 1575-1582, November 2000, Copyright © 2000, The Histochemical Society, Inc.

TECHNICAL NOTE

Methods for Imaging Labeled Neurons Together with Neuropil Features in Drosophila

N. Mark Tyrera, David Shepherdb, and Darren W. Williamsb
a Department of Optometry and Neuroscience, University of Manchester Institute of Science and Technology, Manchester, United Kingdom
b Department of Biology, University of Southampton, Southhampton, United Kingdom

Correspondence to: N. Mark Tyrer, Dept. of Optometry and Neuroscience, U. of Manchester Institute of Science and Technology, Manchester, UK M60 1QD. E-mail: mark.tyrer@umist.ac.uk

We describe staining protocols for serial semithin sections of Drosophila central ganglia that allow visualization of gene expression in particular neurons with counterstaining to display the ganglion architecture. Green fluorescent protein (GFP), expressed in a subset of sensory neurons from a selected enhancer trap line, is visualized by conventional immunohistochemistry with a peroxidase-linked antibody, and neural architecture is revealed by reduced silver staining. This makes visible in histological sections the same GFP-labeled cells seen with confocal microscopy, but with the especial advantage that neuropil structures are also revealed at the level of individual cells and neuron processes. Not only does this allow the physical relationships among intracellularly labeled neurons to be determined by reference to specific features in the neuropil but it also enables a function to be ascribed provisionally to particular regions of neuropil. These methods have particular utility for mapping morphological information on specific neurons in the context of central nervous system architecture, both in adult Drosophila and during development.

(J Histochem Cytochem 48:1575–1581, 2000)

Key Words: Drosophila, central nervous system, semithin sections, serial sectioning, gene expression immunohistochemistry, silver staining, database, three-dimensional reconstruction


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