Cargando…

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...

Descripción completa

Detalles Bibliográficos
Autores principales: Barghi, Neda, Ramirez-Lanzas, Claudia
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Nature Publishing Group UK 2023
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
_version_ 1784902301136191488
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
work_keys_str_mv AT barghineda ahighthroughputmethodforeggsizemeasurementindrosophila
AT ramirezlanzasclaudia ahighthroughputmethodforeggsizemeasurementindrosophila
AT barghineda highthroughputmethodforeggsizemeasurementindrosophila
AT ramirezlanzasclaudia highthroughputmethodforeggsizemeasurementindrosophila