Microscopy techniques in flavivirus research

• A wide range of microscopy techniques from normal light microscopy to super-resolution microscopy and electron microscopy are discussed in detail.
• Various useful F-techniques in microscopy applicable for flavivirus research are reviewed.
• The history and recent advancement of electron microscopy techniques are also discussed with respect to flavivirus research.
• The role of microscopy in flavivirus research is irreplaceable and it will continue to contribute significantly in understanding the life cycle of flavivirus.

The Flavivirus genus is composed of many medically important viruses that cause high morbidity and mortality, which include Dengue and West Nile viruses. Various molecular and biochemical techniques have been developed in the endeavour to study flaviviruses. However, microscopy techniques still have irreplaceable roles in the identification of novel virus pathogens and characterization of morphological changes in virus-infected cells. Fluorescence microscopy contributes greatly in understanding the fundamental viral protein localizations and virus–host protein interactions during infection. Electron microscopy remains the gold standard for visualizing ultra-structural features of virus particles and infected cells. New imaging techniques and combinatory applications are continuously being developed to push the limit of resolution and extract more quantitative data. Currently, correlative live cell imaging and high resolution three-dimensional imaging have already been achieved through the tandem use of optical and electron microscopy in analyzing biological specimens. Microscopy techniques are also used to measure protein binding affinities and determine the mobility pattern of proteins in cells. This chapter will consolidate on the applications of various well-established microscopy techniques in flavivirus research, and discuss how recently developed microscopy techniques can potentially help advance our understanding in these membrane viruses.