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An integrated platform for large-scale data collection and precise perturbation of live Drosophila embryos

Understanding the fundamental principles governing embryogenesis is a key goal of developmental biology. Direct observation of embryogenesis via in vivo live imaging is vital to understanding embryogenesis; yet, tedious sample preparation makes it difficult to acquire large-scale imaging data that i...

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Autores principales: Levario, Thomas J., Zhao, Charles, Rouse, Tel, Shvartsman, Stanislav Y., Lu, Hang
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Nature Publishing Group 2016
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4750044/
https://www.ncbi.nlm.nih.gov/pubmed/26864815
http://dx.doi.org/10.1038/srep21366
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author Levario, Thomas J.
Zhao, Charles
Rouse, Tel
Shvartsman, Stanislav Y.
Lu, Hang
author_facet Levario, Thomas J.
Zhao, Charles
Rouse, Tel
Shvartsman, Stanislav Y.
Lu, Hang
author_sort Levario, Thomas J.
collection PubMed
description Understanding the fundamental principles governing embryogenesis is a key goal of developmental biology. Direct observation of embryogenesis via in vivo live imaging is vital to understanding embryogenesis; yet, tedious sample preparation makes it difficult to acquire large-scale imaging data that is often required to overcome experimental and biological noises for quantitative studies. Furthermore, it is often difficult, and sometimes impossible, to incorporate environmental perturbation for understanding developmental responses to external stimuli. To address this issue, we have developed a method for high-throughput imaging of live embryos, delivering precise environmental perturbations, and unbiased data extraction. This platform includes an optimized microfluidic device specifically for live embryos and also for precise perturbations in the microenvironment of the developing embryos. In addition, we developed software for simple, yet accurate, automated segmentation of fluorescent images, and automated data extraction. Using a quantitative assessment we find that embryos develop normally within the microfluidic device. Finally, we show an application of the high-throughput assay for monitoring developmental responses to external stimuli: anoxia-induced developmental arrest in Drosophila embryos. With slight modifications, the method developed in this work can be applied to many other models of development and other stimulus-response behaviors during development.
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spelling pubmed-47500442016-02-18 An integrated platform for large-scale data collection and precise perturbation of live Drosophila embryos Levario, Thomas J. Zhao, Charles Rouse, Tel Shvartsman, Stanislav Y. Lu, Hang Sci Rep Article Understanding the fundamental principles governing embryogenesis is a key goal of developmental biology. Direct observation of embryogenesis via in vivo live imaging is vital to understanding embryogenesis; yet, tedious sample preparation makes it difficult to acquire large-scale imaging data that is often required to overcome experimental and biological noises for quantitative studies. Furthermore, it is often difficult, and sometimes impossible, to incorporate environmental perturbation for understanding developmental responses to external stimuli. To address this issue, we have developed a method for high-throughput imaging of live embryos, delivering precise environmental perturbations, and unbiased data extraction. This platform includes an optimized microfluidic device specifically for live embryos and also for precise perturbations in the microenvironment of the developing embryos. In addition, we developed software for simple, yet accurate, automated segmentation of fluorescent images, and automated data extraction. Using a quantitative assessment we find that embryos develop normally within the microfluidic device. Finally, we show an application of the high-throughput assay for monitoring developmental responses to external stimuli: anoxia-induced developmental arrest in Drosophila embryos. With slight modifications, the method developed in this work can be applied to many other models of development and other stimulus-response behaviors during development. Nature Publishing Group 2016-02-11 /pmc/articles/PMC4750044/ /pubmed/26864815 http://dx.doi.org/10.1038/srep21366 Text en Copyright © 2016, Macmillan Publishers Limited http://creativecommons.org/licenses/by/4.0/ This work is licensed under a Creative Commons Attribution 4.0 International License. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in the credit line; if the material is not included under the Creative Commons license, users will need to obtain permission from the license holder to reproduce the material. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/
spellingShingle Article
Levario, Thomas J.
Zhao, Charles
Rouse, Tel
Shvartsman, Stanislav Y.
Lu, Hang
An integrated platform for large-scale data collection and precise perturbation of live Drosophila embryos
title An integrated platform for large-scale data collection and precise perturbation of live Drosophila embryos
title_full An integrated platform for large-scale data collection and precise perturbation of live Drosophila embryos
title_fullStr An integrated platform for large-scale data collection and precise perturbation of live Drosophila embryos
title_full_unstemmed An integrated platform for large-scale data collection and precise perturbation of live Drosophila embryos
title_short An integrated platform for large-scale data collection and precise perturbation of live Drosophila embryos
title_sort integrated platform for large-scale data collection and precise perturbation of live drosophila embryos
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4750044/
https://www.ncbi.nlm.nih.gov/pubmed/26864815
http://dx.doi.org/10.1038/srep21366
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