Abstract

During embryogenesis, cells repeatedly divide and dynamically change their positions in three-dimensional (3D) space. A robust and accurate algorithm to acquire the 3D positions of the cells would help to reveal the mechanisms of embryogenesis. To acquire quantitative criteria of embryogenesis from time-series 3D microscopic images, image processing algorithms such as segmentation have been applied. Because the cells in embryos are considerably crowded, an algorithm to segment individual cells in detail and accurately is needed. To quantify the nuclear region of every cell from a time-series 3D fluorescence microscopic image of living cells, we developed QCANet, a convolutional neural network-based segmentation algorithm for 3D fluorescence bioimages. We demonstrated that QCANet outperformed 3D Mask R-CNN, which is currently considered as the best algorithm of instance segmentation. We showed that QCANet can be applied not only to developing mouse embryos but also to developing embryos of two other model species. Using QCANet, we were able to extract several quantitative criteria of embryogenesis from 11 early mouse embryos. We showed that the extracted criteria could be used to evaluate the differences between individual embryos. This study contributes to the development of fundamental approaches for assessing embryogenesis on the basis of extracted quantitative criteria.

Highlights

  • During embryogenesis, cells repeatedly divide and dynamically change their positions in three-dimensional (3D) space[1]

  • The implemented algorithm QCANet is a tool for instance segmentation of 3D fluorescence microscopic images (Fig. 2)

  • QCANet consists of two subnetworks: Nuclear Segmentation Network (NSN) and Nuclear Detection Network (NDN)

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Summary

Introduction

Cells repeatedly divide and dynamically change their positions in three-dimensional (3D) space[1]. A robust and accurate algorithm to acquire the 3D positions of the cells with high temporal resolution would undoubtedly help to reveal the mechanisms of embryogenesis. To analyse the time-series 3D microscopic images of developing embryos with fluorescently labelled nuclei, these studies used image segmentation. Segmentation algorithms in bioimage processing (such as filtering, thresholding, morphological operations, watershed transformation and mask processing7,17–21) require some parameter values. Because these algorithms are based on heuristic image-processing algorithms, they fail to detect an object in an image when this object does not fit the pattern that the algorithm can process. It is hard to accurately acquire quantitative criteria with the existing heuristic image processingbased segmentation algorithms

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