Repository logo
 

Contour, a semi-automated segmentation and quantitation tool for cryo-soft-X-ray tomography

Published version
Peer-reviewed

Type

Article

Change log

Authors

Ferreira Fernandes, João 
Vyas, Nina 

Abstract

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. Segmentation of these compartments is limited by time-consuming manual tools or machine learning algorithms that require extensive time and effort to train. Here we describe Contour, a new, easy-to-use, highly automated segmentation tool that enables accelerated segmentation of tomograms to delineate distinct cellular compartments. 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. 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. 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. Contour can extract geometric measurements from 3D segmented volumes, providing a new method to quantitate cryo-soft-X-ray tomography data. Contour can be freely downloaded at github.com/kamallouisnahas/Contour.

Description

Keywords

3D imaging, cryoSXT, microscopy, quantitation, segmentation

Journal Title

Biological Imaging

Conference Name

Journal ISSN

2633-903X
2633-903X

Volume Title

Publisher

Cambridge University Press
Sponsorship
Biotechnology and Biological Sciences Research Council (BB/M021424/1)
Wellcome Trust (098406/Z/12/B)
Wellcome Trust (098406/Z/12/Z)
Diamond Light Source Ltd.