3/29/2023 0 Comments Easy contour tools(B) In Contour, a width restriction was applied in addition to an intensity threshold to segment the mitochondria. However, intensity thresholding alone led to noise and non-specific features such as the outline of lipid droplets being included in the segmented volume (orange arrows). This technique was highly sensitive as most of the mitochondria were included and only a few areas were missing (white arrows). (A) The mitochondria in a tomogram of a U2OS cell were segmented by applying a voxel intensity threshold (blue arrows) LD, lipid droplet Mito, mitochondrion. Contour can be freely downloaded at /kamallouisnahas/Contour.ģD imaging cryoSXT microscopy quantitation segmentation. Contour can extract geometric measurements from 3D segmented volumes, providing a new method to quantitate cryo-soft-X-ray tomography data. We show that high-contrast compartments such as mitochondria, lipid droplets, and features at the cell surface can be easily segmented with this technique in the context of investigating herpes simplex virus 1 infection. This method is less laborious and less prone to errors from human judgement than current tools that require features to be manually traced, and does not require training datasets as would machine-learning driven segmentation. Using Contour, cellular structures can be segmented based on their projection intensity and geometrical width by applying a threshold range to the image and excluding noise smaller in width than the cellular compartments of interest. Here we describe Contour, a new, easy-to-use, highly automated segmentation tool that enables accelerated segmentation of tomograms to delineate distinct cellular compartments. Segmentation of these compartments is limited by time-consuming manual tools or machine learning algorithms that require extensive time and effort to train. Cryo-soft-X-ray tomography is being increasingly used in biological research to study the morphology of cellular compartments and how they change in response to different stimuli, such as viral infections.
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