Streamlined montage cryo-electron tomography for exploring the ultrastructure of cells and tissues

Cryo-electron tomography reveals the architecture of biological specimens at molecular resolution, albeit with a limited field of view. Montage tomography overcomes this limitation, but existing strategies depend on complex dose distribution imaging schemes or hardware modifications which prevent widespread adoption of the method. To lower the barrier to entry, we developed montaging via streamlined acquisition and integration of cryotomograms (MOSAIC), a straightforward workflow that deviates minimally from conventional image acquisition and leverages our software MontageMaker to merge overlapping tilt series. The resulting montage tomograms are produced with high throughput, retain nanoscale detail despite the uneven accumulation of dose, and are easily acquired on a wide variety of biological samples. Most importantly, this approach can be implemented immediately by any lab with access to automated cryo-ET data collection.