Extended Data Fig. 7: Localization of individual nuclear and cytoplasmic mScarlet-UL25 labelled capsids.
From: Molecular plasticity of herpesvirus nuclear egress analysed in situ
a, PK15 cells were infected with PrV-mScarlet-UL25 or b, PrV-mScarlet-UL25-∆US3 fixed at 7 or 10 hpi, respectively, and imaged using spinning disc microscopy. UL25-mScarlet (red); DNA-Hoechst (blue). One plane of the acquired volume is shown. c, Viral particles were detected in the 3D volumes using Trackmate in FIJI with an expected blob diameter of 0.4 microns and the quality threshold set to 10.0. c, Fluorescent signal in a single plane of a PK15 cell infected with PrV-mScarlet-UL25-∆US3 and fixed 10 hpi (top) and projection of all detected single particles (purple) of the volume in a nuclear ROI (yellow) onto one plane (bottom). d, e, The FIJI plugin Trackmate was used to detect and measure individual virus particle fluorescent intensities. For each condition, the total intensity of more than 4,000 particles was quantified and detected in more than 30 different cells, all using a single biological replicate.
