An open-source semi-automated robotics pipeline for embryo immunohistochemistry

Abstract A significant challenge for developmental systems biology is balancing throughput with controlled conditions that minimize experimental artifacts. Large-scale developmental screens such as unbiased mutagenesis surveys have been limited in their applicability to embryonic systems, as the tec...

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Autores principales:	Timothy Fuqua, Jeff Jordan, Aliaksandr Halavatyi, Christian Tischer, Kerstin Richter, Justin Crocker
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2021
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/9b02fcd58f7a4e718c764ec735d7ce25
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spelling	oai:doaj.org-article:9b02fcd58f7a4e718c764ec735d7ce252021-12-02T16:50:31ZAn open-source semi-automated robotics pipeline for embryo immunohistochemistry10.1038/s41598-021-89676-52045-2322https://doaj.org/article/9b02fcd58f7a4e718c764ec735d7ce252021-05-01T00:00:00Zhttps://doi.org/10.1038/s41598-021-89676-5https://doaj.org/toc/2045-2322Abstract A significant challenge for developmental systems biology is balancing throughput with controlled conditions that minimize experimental artifacts. Large-scale developmental screens such as unbiased mutagenesis surveys have been limited in their applicability to embryonic systems, as the technologies for quantifying precise expression patterns in whole animals has not kept pace with other sequencing-based technologies. Here, we outline an open-source semi-automated pipeline to chemically fixate, stain, and 3D-image Drosophila embryos. Central to this pipeline is a liquid handling robot, Flyspresso, which automates the steps of classical embryo fixation and staining. We provide the schematics and an overview of the technology for an engineer or someone equivalently trained to reproduce and further improve upon Flyspresso, and highlight the Drosophila embryo fixation and colorimetric or antibody staining protocols. Additionally, we provide a detailed overview and stepwise protocol for our adaptive-feedback pipeline for automated embryo imaging on confocal microscopes. We demonstrate the efficiency of this pipeline compared to classical techniques, and how it can be repurposed or scaled to other protocols and biological systems. We hope our pipeline will serve as a platform for future research, allowing a broader community of users to build, execute, and share similar experiments.Timothy FuquaJeff JordanAliaksandr HalavatyiChristian TischerKerstin RichterJustin CrockerNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 11, Iss 1, Pp 1-16 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Timothy Fuqua Jeff Jordan Aliaksandr Halavatyi Christian Tischer Kerstin Richter Justin Crocker An open-source semi-automated robotics pipeline for embryo immunohistochemistry
description	Abstract A significant challenge for developmental systems biology is balancing throughput with controlled conditions that minimize experimental artifacts. Large-scale developmental screens such as unbiased mutagenesis surveys have been limited in their applicability to embryonic systems, as the technologies for quantifying precise expression patterns in whole animals has not kept pace with other sequencing-based technologies. Here, we outline an open-source semi-automated pipeline to chemically fixate, stain, and 3D-image Drosophila embryos. Central to this pipeline is a liquid handling robot, Flyspresso, which automates the steps of classical embryo fixation and staining. We provide the schematics and an overview of the technology for an engineer or someone equivalently trained to reproduce and further improve upon Flyspresso, and highlight the Drosophila embryo fixation and colorimetric or antibody staining protocols. Additionally, we provide a detailed overview and stepwise protocol for our adaptive-feedback pipeline for automated embryo imaging on confocal microscopes. We demonstrate the efficiency of this pipeline compared to classical techniques, and how it can be repurposed or scaled to other protocols and biological systems. We hope our pipeline will serve as a platform for future research, allowing a broader community of users to build, execute, and share similar experiments.
format	article
author	Timothy Fuqua Jeff Jordan Aliaksandr Halavatyi Christian Tischer Kerstin Richter Justin Crocker
author_facet	Timothy Fuqua Jeff Jordan Aliaksandr Halavatyi Christian Tischer Kerstin Richter Justin Crocker
author_sort	Timothy Fuqua
title	An open-source semi-automated robotics pipeline for embryo immunohistochemistry
title_short	An open-source semi-automated robotics pipeline for embryo immunohistochemistry
title_full	An open-source semi-automated robotics pipeline for embryo immunohistochemistry
title_fullStr	An open-source semi-automated robotics pipeline for embryo immunohistochemistry
title_full_unstemmed	An open-source semi-automated robotics pipeline for embryo immunohistochemistry
title_sort	open-source semi-automated robotics pipeline for embryo immunohistochemistry
publisher	Nature Portfolio
publishDate	2021
url	https://doaj.org/article/9b02fcd58f7a4e718c764ec735d7ce25
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An open-source semi-automated robotics pipeline for embryo immunohistochemistry

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