Sigal, Y.M. et al. Cell 163, 493–505 (2015).

Fluorescence microscopy is a valuable tool for volumetric neural circuit reconstruction, but small structures such as synapses can be difficult to identify when imaged at diffraction-limited resolution. To address this issue and image neurons and synapses at high resolution over a large volume, Sigal et al. combined stochastic optical reconstruction microscopy (STORM) with serial ultra-thin sectioning. STORM was used to provide super-resolution images of neurons and synaptic proteins in each section, and the images were then aligned to generate 3D volumetric reconstructions using automated image-analysis tools. The authors used their platform to study on-off direction-selective ganglion cells in the mouse retina and were able to gain new insights into the structural basis of crossover inhibition in these cells.