Confocal quality imaging of afferent neurons from semi-thin sections of Drosophila ganglia.

The aim of this study was to develop protocols for computer imaging of the thoraco-abdominal ganglion of Drosophila from serial semi-thin sections, in which specific neurons were stained and related to neuropilar structures. The central projections of a subset of transgenically labelled sensory neurons were revealed by immunohistochemistry, while Nomarski optics were used to show motor neuron targets in the neuropil. Digital photomicrographs of each section were aligned and the resultant image stacks rendered into three-dimensional (3D) images that can be rotated in real time.

Methods for imaging labeled neurons together with neuropil features in Drosophila.

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.

Celloidin-wax sandwich microtomy: a novel and rapid method for producing serial semithin sections.

A method is described that allows rapid and reliable serial sectioning down to thicknesses of 1 micron. The tissue is first embedded in celloidin and then in wax and trimmed so that the block is sandwiched between two layers of wax. This combines the virtues of both media.

Innervation of dipteran eclosion muscles: ultrastructure, immunohistochemistry, physiology and death.

The thoracic eclosion muscles of flies die by cytotoxic attack under neural control. We have investigated the innervation, ultrastructure and immunohistochemistry of the ventral eclosion muscle of Glossina. Two neurons located in the thoracic ganglion innervate this muscle.