AbstractCryo-electron tomography maps often exhibit considerable noise and anisotropic resolution, due to the low-dose requirements and the missing wedge in Fourier space. These spurious features are visually unappealing and, more importantly, prevent an automated segmentation of geometric shapes, requiring a highly subjective, labor-intensive manual tracing. We developed a novel computational strategy for objectively denoising and correcting missing-wedge artifacts in the special but important case of repetitive basic shapes, such as filamentous structures. In this approach, we use the template and a non-negative “location map” to constrain the deconvolution scheme, allowing us to recover, to a considerable degree, the information lost in the missing wedge. We applied our method to data of actin-filament bundles of inner-ear stereocilia, which are critical in hearing transduction processes, and found a good overlap with the experimental map and with manual tracing. In addition, we demonstrate that our method can also be used for membrane detection.
Rapid low dose electron tomography using a direct electron detection camera