Imaging Flow Cytometry Protocols for Examining Phagocytosis of Microplastics and Bioparticles by Immune Cells of Aquatic Animals
Imaging flow cytometry (IFC) is a powerful tool which combines flow cytometry with digital microscopy to generate quantitative high-throughput imaging data. Despite various advantages of IFC over standard flow cytometry, widespread adoption of this technology for studies in aquatic sciences is limit...
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ftpubmed:oai:pubmedcentral.nih.gov:7039858 2023-05-15T15:31:19+02:00 Imaging Flow Cytometry Protocols for Examining Phagocytosis of Microplastics and Bioparticles by Immune Cells of Aquatic Animals Park, Youngjin Abihssira-García, Isabel S. Thalmann, Sebastian Wiegertjes, Geert F. Barreda, Daniel R. Olsvik, Pål A. Kiron, Viswanath 2020-02-18 http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7039858/ https://doi.org/10.3389/fimmu.2020.00203 en eng Frontiers Media S.A. http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7039858/ http://dx.doi.org/10.3389/fimmu.2020.00203 Copyright © 2020 Park, Abihssira-García, Thalmann, Wiegertjes, Barreda, Olsvik and Kiron. http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms. CC-BY Immunology Text 2020 ftpubmed https://doi.org/10.3389/fimmu.2020.00203 2020-03-08T01:37:42Z Imaging flow cytometry (IFC) is a powerful tool which combines flow cytometry with digital microscopy to generate quantitative high-throughput imaging data. Despite various advantages of IFC over standard flow cytometry, widespread adoption of this technology for studies in aquatic sciences is limited, probably due to the relatively high equipment cost, complexity of image analysis-based data interpretation and lack of core facilities with trained personnel. Here, we describe the application of IFC to examine phagocytosis of particles including microplastics by cells from aquatic animals. For this purpose, we studied (1) live/dead cell assays and identification of cell types, (2) phagocytosis of degradable and non-degradable particles by Atlantic salmon head kidney cells and (3) the effect of incubation temperature on phagocytosis of degradable particles in three aquatic animals–Atlantic salmon, Nile tilapia, and blue mussel. The usefulness of the developed method was assessed by evaluating the effect of incubation temperature on phagocytosis. Our studies demonstrate that IFC provides significant benefits over standard flow cytometry in phagocytosis measurement by allowing integration of morphometric parameters, especially while identifying cell populations and distinguishing between different types of fluorescent particles and detecting their localization. Text Atlantic salmon PubMed Central (PMC) Frontiers in Immunology 11 |
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Immunology Park, Youngjin Abihssira-García, Isabel S. Thalmann, Sebastian Wiegertjes, Geert F. Barreda, Daniel R. Olsvik, Pål A. Kiron, Viswanath Imaging Flow Cytometry Protocols for Examining Phagocytosis of Microplastics and Bioparticles by Immune Cells of Aquatic Animals |
topic_facet |
Immunology |
description |
Imaging flow cytometry (IFC) is a powerful tool which combines flow cytometry with digital microscopy to generate quantitative high-throughput imaging data. Despite various advantages of IFC over standard flow cytometry, widespread adoption of this technology for studies in aquatic sciences is limited, probably due to the relatively high equipment cost, complexity of image analysis-based data interpretation and lack of core facilities with trained personnel. Here, we describe the application of IFC to examine phagocytosis of particles including microplastics by cells from aquatic animals. For this purpose, we studied (1) live/dead cell assays and identification of cell types, (2) phagocytosis of degradable and non-degradable particles by Atlantic salmon head kidney cells and (3) the effect of incubation temperature on phagocytosis of degradable particles in three aquatic animals–Atlantic salmon, Nile tilapia, and blue mussel. The usefulness of the developed method was assessed by evaluating the effect of incubation temperature on phagocytosis. Our studies demonstrate that IFC provides significant benefits over standard flow cytometry in phagocytosis measurement by allowing integration of morphometric parameters, especially while identifying cell populations and distinguishing between different types of fluorescent particles and detecting their localization. |
format |
Text |
author |
Park, Youngjin Abihssira-García, Isabel S. Thalmann, Sebastian Wiegertjes, Geert F. Barreda, Daniel R. Olsvik, Pål A. Kiron, Viswanath |
author_facet |
Park, Youngjin Abihssira-García, Isabel S. Thalmann, Sebastian Wiegertjes, Geert F. Barreda, Daniel R. Olsvik, Pål A. Kiron, Viswanath |
author_sort |
Park, Youngjin |
title |
Imaging Flow Cytometry Protocols for Examining Phagocytosis of Microplastics and Bioparticles by Immune Cells of Aquatic Animals |
title_short |
Imaging Flow Cytometry Protocols for Examining Phagocytosis of Microplastics and Bioparticles by Immune Cells of Aquatic Animals |
title_full |
Imaging Flow Cytometry Protocols for Examining Phagocytosis of Microplastics and Bioparticles by Immune Cells of Aquatic Animals |
title_fullStr |
Imaging Flow Cytometry Protocols for Examining Phagocytosis of Microplastics and Bioparticles by Immune Cells of Aquatic Animals |
title_full_unstemmed |
Imaging Flow Cytometry Protocols for Examining Phagocytosis of Microplastics and Bioparticles by Immune Cells of Aquatic Animals |
title_sort |
imaging flow cytometry protocols for examining phagocytosis of microplastics and bioparticles by immune cells of aquatic animals |
publisher |
Frontiers Media S.A. |
publishDate |
2020 |
url |
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7039858/ https://doi.org/10.3389/fimmu.2020.00203 |
genre |
Atlantic salmon |
genre_facet |
Atlantic salmon |
op_relation |
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7039858/ http://dx.doi.org/10.3389/fimmu.2020.00203 |
op_rights |
Copyright © 2020 Park, Abihssira-García, Thalmann, Wiegertjes, Barreda, Olsvik and Kiron. http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms. |
op_rightsnorm |
CC-BY |
op_doi |
https://doi.org/10.3389/fimmu.2020.00203 |
container_title |
Frontiers in Immunology |
container_volume |
11 |
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1766361807681748992 |