In addition, astigmatism has been used with two-color STORM for 3D SR imaging together with EM and STED to determine the spatial distribution of proteins EphB2 and SynCAM in relation to the postsynaptic density, which revealed that SynCAM 1 shapes the cleft edge, while EphB2 is enriched deeper into the postsynapse (Perez de Arce et al., 2015), as well as for imaging of the distribution of presynaptic proteins in relation to the postsynaptic scaffolding protein PSD-95, which revealed trans-synaptic alignment of the distributions (Tang et al., 2016; Figure 5D). Here, EPHB2 is linked to Astigmatism.