Spatiotemporal dynamics of HSV genome nuclear entry and compaction state transitions using bioorthogonal chemistry and super-resolution microscopy
Equivalent samples of HSV-1EdC or HSV-1 at 1x108 pfu/ml were adsorbed onto glass coverslips prior to detection by cycloaddition and immunofluorescence for VP5. Panels I and IV show the merged channel images for each virus (scale bar 10 μm). The inset in panel I shows a magnified section. Panels II and V show only the green channel (genome detection) for each virus. Panels III and VI show the colour-coded outline overlay produced by the ImageJ plugin used for particle analysis (described in materials and methods); yellow indicates particles containing both VP5 capsid protein and EdC genome signal; red indicates VP5+ve particles lacking detectable EdC; green indicates particles with detectable EdC but no VP5. The data for approximately 700 particles are quantified in the right panels for each virus.