Preparation of Herpes Simplex Virus-Infected Primary Neurons for Transmission Electron Microscopy
Transmission electron microscopy (TEM) provides the resolution necessary to identify both viruses and subcellular components of cells infected with many types of viruses, including herpes simplex virus. Recognized as a powerful tool in both diagnostic and research-based virology laboratories, TEM has made possible the identification of new viruses and has contributed to the elucidation of virus life cycle and virus–host cell interaction.
While there are many sample preparation techniques for TEM, conventional processing using chemical fixation and resin embedding remains a useful technique, available in virtually all EM laboratories, for studying virus/cell ultrastructure. In this chapter, we describe the preparation of herpes simplex virus infected primary neurons, grown on plastic coverslips, to allow for sectioning of neurons and axons in their growth plane. This technique allows for TEM examination of cell bodies, axons, growth cones and varicosities, providing powerful insights into virus–cell interaction.
Key wordsTransmission electron microscopy Neurons Herpes simplex virus Axons Growth cones
This work was supported by the Australian National Health and Medical Research Grants (402457 and 570849), the Westmead Millennium Institute, and the Westmead Medical Research Foundation.
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