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      Automated cellular annotation for high-resolution images of adult Caenorhabditis elegans

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          Abstract

          Motivation: Advances in high-resolution microscopy have recently made possible the analysis of gene expression at the level of individual cells. The fixed lineage of cells in the adult worm Caenorhabditis elegans makes this organism an ideal model for studying complex biological processes like development and aging. However, annotating individual cells in images of adult C.elegans typically requires expertise and significant manual effort. Automation of this task is therefore critical to enabling high-resolution studies of a large number of genes.

          Results: In this article, we describe an automated method for annotating a subset of 154 cells (including various muscle, intestinal and hypodermal cells) in high-resolution images of adult C.elegans. We formulate the task of labeling cells within an image as a combinatorial optimization problem, where the goal is to minimize a scoring function that compares cells in a test input image with cells from a training atlas of manually annotated worms according to various spatial and morphological characteristics. We propose an approach for solving this problem based on reduction to minimum-cost maximum-flow and apply a cross-entropy–based learning algorithm to tune the weights of our scoring function. We achieve 84% median accuracy across a set of 154 cell labels in this highly variable system. These results demonstrate the feasibility of the automatic annotation of microscopy-based images in adult C.elegans.

          Contact: saerni@ 123456cs.stanford.edu

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          Most cited references27

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          Theoretical Improvements in Algorithmic Efficiency for Network Flow Problems

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            Automated cell lineage tracing in Caenorhabditis elegans.

            The invariant cell lineage and cell fate of Caenorhabditis elegans provide a unique opportunity to decode the molecular mechanisms of animal development. To exploit this opportunity, we have developed a system for automated cell lineage tracing during C. elegans embryogenesis, based on 3D, time-lapse imaging and automated image analysis. Using ubiquitously expressed histone-GFP fusion protein to label cells/nuclei and a confocal microscope, the imaging protocol captures embryogenesis at high spatial (31 planes at 1 microm apart) and temporal (every minute) resolution without apparent effects on development. A set of image analysis algorithms then automatically recognizes cells at each time point, tracks cell movements, divisions and deaths over time and assigns cell identities based on the canonical naming scheme. Starting from the four-cell stage (or earlier), our software, named starrynite, can trace the lineage up to the 350-cell stage in 25 min on a desktop computer. The few errors of automated lineaging can then be corrected in a few hours with a graphic interface that allows easy navigation of the images and the reported lineage tree. The system can be used to characterize lineage phenotypes of genes and/or extended to determine gene expression patterns in a living embryo at the single-cell level. We envision that this automation will make it practical to systematically decipher the developmental genes and pathways encoded in the genome of C. elegans.
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              A 3D Digital Atlas of C. elegans and Its Application To Single-Cell Analyses

              We have built a digital nuclear atlas of the newly hatched, first larval stage (L1) of the wild type hermaphrodite of C. elegans at single cell resolution from confocal image stacks of 15 individuals. The atlas quantifies the stereotypy of the locations and provides for other statistics on the spatial patterns of the 357 nuclei that could be faithfully segmented and annotated of the 558 present at this developmental stage. Given this atlas we then developed an automated approach to assign cell names to each nucleus in a 3D image of an L1 worm. We achieve 86% accuracy in identifying the 357 nuclei automatically. This computational method is essential for high-throughput single cell analyses of the worm at post-embryonic stages, such as determining the expression of every gene in every cell during development from the L1 onward, or ablating or stimulating cells under computer control in a high-throughput functional screen.
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                Author and article information

                Journal
                Bioinformatics
                Bioinformatics
                bioinformatics
                bioinfo
                Bioinformatics
                Oxford University Press
                1367-4803
                1367-4811
                1 July 2013
                19 June 2013
                19 June 2013
                : 29
                : 13
                : i18-i26
                Affiliations
                1Department of Computer Science, Stanford University, 353 Serra Mall, Stanford, CA 94305, USA, 2Biomedical Informatics Training Program, Stanford University School of Medicine, 251 Campus Drive, Stanford, CA 94305, USA, 3School of Life Sciences, Tsinghua University, Beijing 100084, China, 423andMe, Inc., 1390 Shorebird Way Mountain View, CA 94043, USA, 5Department of Developmental Biology, Stanford University School of Medicine, 279 Campus Drive Beckman Center B300, Stanford, CA 94305-5329, USA, 6Janelia Farm Research Campus, Howard Hughes Medical Institute, Ashburn, VA, USA and 7Allen Institute for Brain Science, Seattle, WA, USA
                Author notes
                *To whom correspondence should be addressed.
                Article
                btt223
                10.1093/bioinformatics/btt223
                3694659
                23812982
                934bcd26-0cc7-4f30-97ea-ac56ae76350d
                © The Author 2013. Published by Oxford University Press.

                This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License ( http://creativecommons.org/licenses/by-nc/3.0/), which permits non-commercial re-use, distribution, and reproduction in any medium, provided the original work is properly cited. For commercial re-use, please contact journals.permissions@oup.com

                History
                Page count
                Pages: 9
                Categories
                Ismb/Eccb 2013 Proceedings Papers Committee July 21 to July 23, 2013, Berlin, Germany
                Original Papers
                Bioimaging and Data Visualization

                Bioinformatics & Computational biology
                Bioinformatics & Computational biology

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