Conventional, High-Resolution and Imaging Flow Cytometry: Benchmarking Performance in Characterisation of Extracellular Vesicles
Flow cytometry remains a commonly used methodology due to its ability to characterise multiple parameters on single particles in a high-throughput manner. In order to address limitations with lacking sensitivity of conventional flow cytometry to characterise extracellular vesicles (EVs), novel, high...
| Main Authors: | , , |
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| Format: | Article |
| Language: | English |
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MDPI AG
2021-01-01
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| Series: | Biomedicines |
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| Online Access: | https://www.mdpi.com/2227-9059/9/2/124 |
| _version_ | 1797406923385995264 |
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| author | Jaco Botha Haley R. Pugsley Aase Handberg |
| author_facet | Jaco Botha Haley R. Pugsley Aase Handberg |
| author_sort | Jaco Botha |
| collection | DOAJ |
| description | Flow cytometry remains a commonly used methodology due to its ability to characterise multiple parameters on single particles in a high-throughput manner. In order to address limitations with lacking sensitivity of conventional flow cytometry to characterise extracellular vesicles (EVs), novel, highly sensitive platforms, such as high-resolution and imaging flow cytometers, have been developed. We provided comparative benchmarks of a conventional FACS Aria III, a high-resolution Apogee A60 Micro-PLUS and the ImageStream X Mk II imaging flow cytometry platform. Nanospheres were used to systematically characterise the abilities of each platform to detect and quantify populations with different sizes, refractive indices and fluorescence properties, and the repeatability in concentration determinations was reported for each population. We evaluated the ability of the three platforms to detect different EV phenotypes in blood plasma and the intra-day, inter-day and global variabilities in determining EV concentrations. By applying this or similar methodology to characterise methods, researchers would be able to make informed decisions on choice of platforms and thereby be able to match suitable flow cytometry platforms with projects based on the needs of each individual project. This would greatly contribute to improving the robustness and reproducibility of EV studies. |
| first_indexed | 2024-03-09T03:33:47Z |
| format | Article |
| id | doaj.art-aac94592029d4a37b0b93d6b745fd9c8 |
| institution | Directory Open Access Journal |
| issn | 2227-9059 |
| language | English |
| last_indexed | 2024-03-09T03:33:47Z |
| publishDate | 2021-01-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Biomedicines |
| spelling | doaj.art-aac94592029d4a37b0b93d6b745fd9c82023-12-03T14:52:22ZengMDPI AGBiomedicines2227-90592021-01-019212410.3390/biomedicines9020124Conventional, High-Resolution and Imaging Flow Cytometry: Benchmarking Performance in Characterisation of Extracellular VesiclesJaco Botha0Haley R. Pugsley1Aase Handberg2Department of Clinical Biochemistry, Aalborg University Hospital, North Denmark Region, DK-9000 Aalborg, DenmarkLuminex Corporation, Seattle, WA 98119, USADepartment of Clinical Biochemistry, Aalborg University Hospital, North Denmark Region, DK-9000 Aalborg, DenmarkFlow cytometry remains a commonly used methodology due to its ability to characterise multiple parameters on single particles in a high-throughput manner. In order to address limitations with lacking sensitivity of conventional flow cytometry to characterise extracellular vesicles (EVs), novel, highly sensitive platforms, such as high-resolution and imaging flow cytometers, have been developed. We provided comparative benchmarks of a conventional FACS Aria III, a high-resolution Apogee A60 Micro-PLUS and the ImageStream X Mk II imaging flow cytometry platform. Nanospheres were used to systematically characterise the abilities of each platform to detect and quantify populations with different sizes, refractive indices and fluorescence properties, and the repeatability in concentration determinations was reported for each population. We evaluated the ability of the three platforms to detect different EV phenotypes in blood plasma and the intra-day, inter-day and global variabilities in determining EV concentrations. By applying this or similar methodology to characterise methods, researchers would be able to make informed decisions on choice of platforms and thereby be able to match suitable flow cytometry platforms with projects based on the needs of each individual project. This would greatly contribute to improving the robustness and reproducibility of EV studies.https://www.mdpi.com/2227-9059/9/2/124extracellular vesiclesexosomesmicrovesiclesflow cytometryimaging flow cytometryhigh-resolution flow cytometry |
| spellingShingle | Jaco Botha Haley R. Pugsley Aase Handberg Conventional, High-Resolution and Imaging Flow Cytometry: Benchmarking Performance in Characterisation of Extracellular Vesicles Biomedicines extracellular vesicles exosomes microvesicles flow cytometry imaging flow cytometry high-resolution flow cytometry |
| title | Conventional, High-Resolution and Imaging Flow Cytometry: Benchmarking Performance in Characterisation of Extracellular Vesicles |
| title_full | Conventional, High-Resolution and Imaging Flow Cytometry: Benchmarking Performance in Characterisation of Extracellular Vesicles |
| title_fullStr | Conventional, High-Resolution and Imaging Flow Cytometry: Benchmarking Performance in Characterisation of Extracellular Vesicles |
| title_full_unstemmed | Conventional, High-Resolution and Imaging Flow Cytometry: Benchmarking Performance in Characterisation of Extracellular Vesicles |
| title_short | Conventional, High-Resolution and Imaging Flow Cytometry: Benchmarking Performance in Characterisation of Extracellular Vesicles |
| title_sort | conventional high resolution and imaging flow cytometry benchmarking performance in characterisation of extracellular vesicles |
| topic | extracellular vesicles exosomes microvesicles flow cytometry imaging flow cytometry high-resolution flow cytometry |
| url | https://www.mdpi.com/2227-9059/9/2/124 |
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