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.

We have demonstrated the use of 2D dSTORM and particle averaging for the precise determination of 3D protein radial distribution in virus particles. In principle, 3D SMLM imaging techniques can also be used, such as dSTORM with astigmatic 55 or biplanar 56 detection schemes. Our method retain the high lateral resolution of 2D dSTORM techniques while recovering 3D information by taking into account the spherical symmetry of the particle. The same approach is applicable for any spherical system, such as other complex viruses, including coronaviruses (for example, the SARS, severe acute respiratory syndrome, or MERS, Middle East respiratory syndrome, coronaviruses) and rotaviruses. However, the spherical symmetry is not an a priori constraint: any system is amenable to the methods presented here as long as structural information is available for accurate modelling of the SMLM data. For more complex structures, the alignment may require more complex algorithms, such as those reported for the alignment of nuclear pore complexes 34 . Another powerful approach to consider would be correlative imaging, combining optical fluorescence microscopy and EM as demonstrated previously 57 . This approach would potentially reveal structural information (notably on the capsid) to complement the molecule-specific fluorescence imaging.