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A high throughput method for egg size measurement in Drosophila
Life-history traits are used as proxies of fitness in insects including Drosophila. Egg size is an adaptive and ecologically important trait potentially with genetic variation across different populations. However, the low throughput of manual measurement of egg size has hampered the widespread use...
Autores principales: | , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Nature Publishing Group UK
2023
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9992389/ https://www.ncbi.nlm.nih.gov/pubmed/36882448 http://dx.doi.org/10.1038/s41598-023-30472-8 |
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author | Barghi, Neda Ramirez-Lanzas, Claudia |
author_facet | Barghi, Neda Ramirez-Lanzas, Claudia |
author_sort | Barghi, Neda |
collection | PubMed |
description | Life-history traits are used as proxies of fitness in insects including Drosophila. Egg size is an adaptive and ecologically important trait potentially with genetic variation across different populations. However, the low throughput of manual measurement of egg size has hampered the widespread use of this trait in evolutionary biology and population genetics. We established a method for accurate and high throughput measurement of Drosophila egg size using large particle flow cytometry (LPFC). The size estimates using LPFC are accurate and highly correlated with the manual measurements. The measurement of egg size is high throughput (average of 214 eggs measured per minute) and viable eggs of a specific size can be sorted rapidly (average of 70 eggs per minute). Sorting by LPFC does not reduce the survival of eggs making it a suitable approach for sorting eggs for downstream analyses. This protocol can be applied to any organism within the detectable size range (10–1500 µm) of the large particle flow cytometers. We discuss the potential applications of this method and provide recommendations for optimizing the protocol for other organisms. |
format | Online Article Text |
id | pubmed-9992389 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2023 |
publisher | Nature Publishing Group UK |
record_format | MEDLINE/PubMed |
spelling | pubmed-99923892023-03-09 A high throughput method for egg size measurement in Drosophila Barghi, Neda Ramirez-Lanzas, Claudia Sci Rep Article Life-history traits are used as proxies of fitness in insects including Drosophila. Egg size is an adaptive and ecologically important trait potentially with genetic variation across different populations. However, the low throughput of manual measurement of egg size has hampered the widespread use of this trait in evolutionary biology and population genetics. We established a method for accurate and high throughput measurement of Drosophila egg size using large particle flow cytometry (LPFC). The size estimates using LPFC are accurate and highly correlated with the manual measurements. The measurement of egg size is high throughput (average of 214 eggs measured per minute) and viable eggs of a specific size can be sorted rapidly (average of 70 eggs per minute). Sorting by LPFC does not reduce the survival of eggs making it a suitable approach for sorting eggs for downstream analyses. This protocol can be applied to any organism within the detectable size range (10–1500 µm) of the large particle flow cytometers. We discuss the potential applications of this method and provide recommendations for optimizing the protocol for other organisms. Nature Publishing Group UK 2023-03-07 /pmc/articles/PMC9992389/ /pubmed/36882448 http://dx.doi.org/10.1038/s41598-023-30472-8 Text en © The Author(s) 2023 https://creativecommons.org/licenses/by/4.0/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 licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence 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 licence, visit http://creativecommons.org/licenses/by/4.0/ (https://creativecommons.org/licenses/by/4.0/) . |
spellingShingle | Article Barghi, Neda Ramirez-Lanzas, Claudia A high throughput method for egg size measurement in Drosophila |
title | A high throughput method for egg size measurement in Drosophila |
title_full | A high throughput method for egg size measurement in Drosophila |
title_fullStr | A high throughput method for egg size measurement in Drosophila |
title_full_unstemmed | A high throughput method for egg size measurement in Drosophila |
title_short | A high throughput method for egg size measurement in Drosophila |
title_sort | high throughput method for egg size measurement in drosophila |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9992389/ https://www.ncbi.nlm.nih.gov/pubmed/36882448 http://dx.doi.org/10.1038/s41598-023-30472-8 |
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