Structural analysis of herpes simplex virus by optical super-resolution imaging

Laine, Romain F.; Albecka, Anna; van de Linde, Sebastian; Rees, Eric J.; Crump, Colin M.; Kaminski, Clemens F.

Here we present a study of HSV-1 viral structure using dSTORM. First, multi-colour localization microscopy was used to visually distinguish virus proteins in individual virus particles. We then designed a model-based analysis of SMLM data and used particle averaging (a method routinely used in EM 34 ) to reconstruct a high-resolution image and to determine the position of individual protein layers within the virus particle with nanometre precision. We additionally investigated the effect of the fluorophore linker size and compared immuno-labelling strategies in the context of model-based analysis. Finally, our data set confirmed and quantified the spatial offset of the capsid with respect to the centre of the virion.