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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| Format: | Article |
| Language: | English |
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Frontiers Media S.A.
2020-02-01
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| Series: | Frontiers in Immunology |
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| Online Access: | https://www.frontiersin.org/article/10.3389/fimmu.2020.00203/full |
| _version_ | 1819082527015436288 |
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| author | Youngjin Park Isabel S. Abihssira-García Sebastian Thalmann Geert F. Wiegertjes Daniel R. Barreda Pål A. Olsvik Viswanath Kiron |
| author_facet | Youngjin Park Isabel S. Abihssira-García Sebastian Thalmann Geert F. Wiegertjes Daniel R. Barreda Pål A. Olsvik Viswanath Kiron |
| author_sort | Youngjin Park |
| collection | DOAJ |
| 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. |
| first_indexed | 2024-12-21T20:18:05Z |
| format | Article |
| id | doaj.art-fc7c99a80bbc4a71810eb3a67ff9f154 |
| institution | Directory Open Access Journal |
| issn | 1664-3224 |
| language | English |
| last_indexed | 2024-12-21T20:18:05Z |
| publishDate | 2020-02-01 |
| publisher | Frontiers Media S.A. |
| record_format | Article |
| series | Frontiers in Immunology |
| spelling | doaj.art-fc7c99a80bbc4a71810eb3a67ff9f1542022-12-21T18:51:34ZengFrontiers Media S.A.Frontiers in Immunology1664-32242020-02-011110.3389/fimmu.2020.00203506168Imaging Flow Cytometry Protocols for Examining Phagocytosis of Microplastics and Bioparticles by Immune Cells of Aquatic AnimalsYoungjin Park0Isabel S. Abihssira-García1Sebastian Thalmann2Geert F. Wiegertjes3Daniel R. Barreda4Pål A. Olsvik5Viswanath Kiron6Faculty of Biosciences and Aquaculture, Nord University, Bodø, NorwayFaculty of Biosciences and Aquaculture, Nord University, Bodø, NorwayLuminex B.V., ‘s-Hertogenbosch, NetherlandsAquaculture and Fisheries Group, Wageningen University & Research, Wageningen, NetherlandsDepartment of Biological Sciences, University of Alberta, Edmonton, AB, CanadaFaculty of Biosciences and Aquaculture, Nord University, Bodø, NorwayFaculty of Biosciences and Aquaculture, Nord University, Bodø, NorwayImaging 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.https://www.frontiersin.org/article/10.3389/fimmu.2020.00203/fullImageStream®XIFCAtlantic salmonNile tilapiablue musselphagocytosis |
| spellingShingle | Youngjin Park Isabel S. Abihssira-García Sebastian Thalmann Geert F. Wiegertjes Daniel R. Barreda Pål A. Olsvik Viswanath Kiron Imaging Flow Cytometry Protocols for Examining Phagocytosis of Microplastics and Bioparticles by Immune Cells of Aquatic Animals Frontiers in Immunology ImageStream®X IFC Atlantic salmon Nile tilapia blue mussel phagocytosis |
| title | 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_short | 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 |
| topic | ImageStream®X IFC Atlantic salmon Nile tilapia blue mussel phagocytosis |
| url | https://www.frontiersin.org/article/10.3389/fimmu.2020.00203/full |
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