Staining and resin embedding of whole Daphnia magna samples for micro-CT imaging enabling 3D visualization of cells, tissues, and organs.

Micro-CT imaging is a powerful tool for generating high-resolution, isotropic, three-dimensional datasets of whole, centimeter-scale model organisms. At histological resolutions, micro-CT can be used for whole-animal qualitative and quantitative characterization of tissue and organismal structure in...

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Main Authors: Mee S Ngu, Daniel J Vanselow, Andrew L Sugarman, Rachelle A Saint-Fort, Carolyn R Zaino, Maksim A Yakovlev, Keith C Cheng, Khai C Ang
Format: Article
Language:English
Published: Public Library of Science (PLoS) 2024-01-01
Series:PLoS ONE
Online Access:https://doi.org/10.1371/journal.pone.0313389
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author Mee S Ngu
Daniel J Vanselow
Andrew L Sugarman
Rachelle A Saint-Fort
Carolyn R Zaino
Maksim A Yakovlev
Keith C Cheng
Khai C Ang
author_facet Mee S Ngu
Daniel J Vanselow
Andrew L Sugarman
Rachelle A Saint-Fort
Carolyn R Zaino
Maksim A Yakovlev
Keith C Cheng
Khai C Ang
author_sort Mee S Ngu
collection DOAJ
description Micro-CT imaging is a powerful tool for generating high-resolution, isotropic, three-dimensional datasets of whole, centimeter-scale model organisms. At histological resolutions, micro-CT can be used for whole-animal qualitative and quantitative characterization of tissue and organismal structure in health and disease. The small size, global freshwater distribution, wide range of cell size and structures of micron scale, and common use of Daphnia magna in toxicological and environmental studies make it an ideal model for demonstrating the potential power of micro-CT-enabled whole-organism phenotyping. This protocol details the steps involved in D. magna samples preparation for micro-CT, including euthanasia, fixation, staining, and resin embedding. Micro-CT reconstructions of samples imaged using synchrotron micro-CT reveal histological (microanatomic) features of organ systems, tissues, and cells in the context of the entire organism at sub-micron resolution and in 3D. The enabled "3D histology" and 3D renderings can be used for morphometric analyses across cells, tissues, and organ systems.
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spelling doaj-art-a8bef96cebb14e3d82f5bb295608e1c72025-08-20T02:41:42ZengPublic Library of Science (PLoS)PLoS ONE1932-62032024-01-011911e031338910.1371/journal.pone.0313389Staining and resin embedding of whole Daphnia magna samples for micro-CT imaging enabling 3D visualization of cells, tissues, and organs.Mee S NguDaniel J VanselowAndrew L SugarmanRachelle A Saint-FortCarolyn R ZainoMaksim A YakovlevKeith C ChengKhai C AngMicro-CT imaging is a powerful tool for generating high-resolution, isotropic, three-dimensional datasets of whole, centimeter-scale model organisms. At histological resolutions, micro-CT can be used for whole-animal qualitative and quantitative characterization of tissue and organismal structure in health and disease. The small size, global freshwater distribution, wide range of cell size and structures of micron scale, and common use of Daphnia magna in toxicological and environmental studies make it an ideal model for demonstrating the potential power of micro-CT-enabled whole-organism phenotyping. This protocol details the steps involved in D. magna samples preparation for micro-CT, including euthanasia, fixation, staining, and resin embedding. Micro-CT reconstructions of samples imaged using synchrotron micro-CT reveal histological (microanatomic) features of organ systems, tissues, and cells in the context of the entire organism at sub-micron resolution and in 3D. The enabled "3D histology" and 3D renderings can be used for morphometric analyses across cells, tissues, and organ systems.https://doi.org/10.1371/journal.pone.0313389
spellingShingle Mee S Ngu
Daniel J Vanselow
Andrew L Sugarman
Rachelle A Saint-Fort
Carolyn R Zaino
Maksim A Yakovlev
Keith C Cheng
Khai C Ang
Staining and resin embedding of whole Daphnia magna samples for micro-CT imaging enabling 3D visualization of cells, tissues, and organs.
PLoS ONE
title Staining and resin embedding of whole Daphnia magna samples for micro-CT imaging enabling 3D visualization of cells, tissues, and organs.
title_full Staining and resin embedding of whole Daphnia magna samples for micro-CT imaging enabling 3D visualization of cells, tissues, and organs.
title_fullStr Staining and resin embedding of whole Daphnia magna samples for micro-CT imaging enabling 3D visualization of cells, tissues, and organs.
title_full_unstemmed Staining and resin embedding of whole Daphnia magna samples for micro-CT imaging enabling 3D visualization of cells, tissues, and organs.
title_short Staining and resin embedding of whole Daphnia magna samples for micro-CT imaging enabling 3D visualization of cells, tissues, and organs.
title_sort staining and resin embedding of whole daphnia magna samples for micro ct imaging enabling 3d visualization of cells tissues and organs
url https://doi.org/10.1371/journal.pone.0313389
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