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Automated, high-throughput measurement of size and growth curves of small organisms in well plates

Organism size and growth curves are important biological characteristics. Current methods to measure organism size, and in particular growth curves, are often resource intensive because they involve many manual steps. Here we demonstrate a method for automated, high-throughput measurements of size a...

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Autores principales: Duckworth, James, Jager, Tjalling, Ashauer, Roman
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Nature Publishing Group UK 2019
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6327043/
https://www.ncbi.nlm.nih.gov/pubmed/30626881
http://dx.doi.org/10.1038/s41598-018-36877-0
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author Duckworth, James
Jager, Tjalling
Ashauer, Roman
author_facet Duckworth, James
Jager, Tjalling
Ashauer, Roman
author_sort Duckworth, James
collection PubMed
description Organism size and growth curves are important biological characteristics. Current methods to measure organism size, and in particular growth curves, are often resource intensive because they involve many manual steps. Here we demonstrate a method for automated, high-throughput measurements of size and growth in individual aquatic invertebrates kept in microtiter well-plates. We use a spheroid counter (Cell(3)iMager, cc-5000) to automatically measure size of seven different freshwater invertebrate species. Further, we generated calibration curves (linear regressions, all p < 0.0001, r(2) >=0.9 for Ceriodaphnoa dubia, Asellus aquaticus, Daphnia magna, Daphnia pulex; r(2) >=0.8 for Hyalella azteca, Chironomus spec. larvae and Culex spec. larvae) to convert size measured on the spheroid counter to traditional, microscope based, length measurements, which follow the longest orientation of the body. Finally, we demonstrate semi-automated measurement of growth curves of individual daphnids (C. dubia and D. magna) over time and find that the quality of individual growth curves varies, partly due to methodological reasons. Nevertheless, this novel method could be adopted to other species and represents a step change in experimental throughput for measuring organisms’ shape, size and growth curves. It is also a significant qualitative improvement by enabling high-throughput assessment of inter-individual variation of growth.
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spelling pubmed-63270432019-01-11 Automated, high-throughput measurement of size and growth curves of small organisms in well plates Duckworth, James Jager, Tjalling Ashauer, Roman Sci Rep Article Organism size and growth curves are important biological characteristics. Current methods to measure organism size, and in particular growth curves, are often resource intensive because they involve many manual steps. Here we demonstrate a method for automated, high-throughput measurements of size and growth in individual aquatic invertebrates kept in microtiter well-plates. We use a spheroid counter (Cell(3)iMager, cc-5000) to automatically measure size of seven different freshwater invertebrate species. Further, we generated calibration curves (linear regressions, all p < 0.0001, r(2) >=0.9 for Ceriodaphnoa dubia, Asellus aquaticus, Daphnia magna, Daphnia pulex; r(2) >=0.8 for Hyalella azteca, Chironomus spec. larvae and Culex spec. larvae) to convert size measured on the spheroid counter to traditional, microscope based, length measurements, which follow the longest orientation of the body. Finally, we demonstrate semi-automated measurement of growth curves of individual daphnids (C. dubia and D. magna) over time and find that the quality of individual growth curves varies, partly due to methodological reasons. Nevertheless, this novel method could be adopted to other species and represents a step change in experimental throughput for measuring organisms’ shape, size and growth curves. It is also a significant qualitative improvement by enabling high-throughput assessment of inter-individual variation of growth. Nature Publishing Group UK 2019-01-09 /pmc/articles/PMC6327043/ /pubmed/30626881 http://dx.doi.org/10.1038/s41598-018-36877-0 Text en © The Author(s) 2019 Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.
spellingShingle Article
Duckworth, James
Jager, Tjalling
Ashauer, Roman
Automated, high-throughput measurement of size and growth curves of small organisms in well plates
title Automated, high-throughput measurement of size and growth curves of small organisms in well plates
title_full Automated, high-throughput measurement of size and growth curves of small organisms in well plates
title_fullStr Automated, high-throughput measurement of size and growth curves of small organisms in well plates
title_full_unstemmed Automated, high-throughput measurement of size and growth curves of small organisms in well plates
title_short Automated, high-throughput measurement of size and growth curves of small organisms in well plates
title_sort automated, high-throughput measurement of size and growth curves of small organisms in well plates
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6327043/
https://www.ncbi.nlm.nih.gov/pubmed/30626881
http://dx.doi.org/10.1038/s41598-018-36877-0
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