Significant strategies for the successful performance of cryo-electron tomography

This technical note describes the preparation of frozen-hydrated specimens and critical imaging conditions for the successful acquisition of data for cryo-electron tomography. Cryo-fixation with appropriate parameters for the plunge-freezing method could aid in the preparation of improved biological cryo-samples. This note then describes in detail several significant constraints for data collection, such as total tolerable electron dose for the acquisition of tilt series images, enhancement of the image contrast, and the need for low electron dose imaging. Together, these strategies were applied for cryo-electron tomographic three-dimensional visualization of the cellular component, multilamella body from a HeLa cell.