Confocal Imaging of Nerve Cells and Their Connections
This chapter outlines the use of confocal microscopy combined with immunofluorescence staining for examining nerve cells in the central nervous system (CNS) and studying the synaptic connections between them. A method for carrying out immunolabeling with two or three different fluorescent dyes is described, together with modifications that allow this to be combined with tract tracing,intracellular injection, or lectin labeling, and also a new technique for combining confocal and electron microscopy (EM) on sections which have been processed for immunocytochemistry. With these approaches, it is possible to search for and quantify contacts between neurons at the light microscopic level and then examine representative contacts at the ultrastructural level to determine whether synapses are present.
KeywordsDorsal Horn Confocal Image Dendritic Tree Primary Afferents Immunoreactive Neuron
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