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Content Size-Dependent Alginate Microcapsule Formation Using Centrifugation to Eliminate Empty Microcapsules for On-Chip Imaging Cell Sorter Application

Alginate microcapsules are one of the attractive non-invasive platforms for handling individual cells and clusters, maintaining their isolation for further applications such as imaging cell sorter and single capsule qPCR. However, the conventional cell encapsulation techniques provide huge numbers o...

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Autores principales: Akimoto, Toshinosuke, Yasuda, Kenji
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9867324/
https://www.ncbi.nlm.nih.gov/pubmed/36677133
http://dx.doi.org/10.3390/mi14010072
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author Akimoto, Toshinosuke
Yasuda, Kenji
author_facet Akimoto, Toshinosuke
Yasuda, Kenji
author_sort Akimoto, Toshinosuke
collection PubMed
description Alginate microcapsules are one of the attractive non-invasive platforms for handling individual cells and clusters, maintaining their isolation for further applications such as imaging cell sorter and single capsule qPCR. However, the conventional cell encapsulation techniques provide huge numbers of unnecessary empty homogeneous alginate microcapsules, which spend an excessive majority of the machine time on observations and analysis. Here, we developed a simple alginate cell encapsulation method to form content size-dependent alginate microcapsules to eliminate empty microcapsules using microcapillary centrifugation and filtration. Using this method, the formed calcium alginate microcapsules containing the HeLa cells were larger than 20m, and the other empty microcapsules were less than 3m under 4000 rpm centrifugation condition. We collected cell-containing alginate microcapsules by eliminating empty microcapsules from the microcapsule mixture with simple one-step filtration of a 20 m cell strainer. The electrical surface charge density and optical permeability of those cell-encapsulated alginate microcapsules were also evaluated. We found that the surface charge density of cell-encapsulated alginate microbeads is more than double that of cells, indicating that less voltage is required for electrical cell handling with thin alginate gel encapsulation of samples. The permeability of the alginate microcapsule was not improved by changing the reflective index of the medium buffer, such as adding alginate ester. However, the minimized thickness of the alginate gel envelope surrounding cells in the microcapsules did not degrade the detailed shapes of encapsulated cells. Those results confirmed the advantage of alginate encapsulation of cells with the centrifugation method as one of the desirable tools for imaging cell sorting applications.
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spelling pubmed-98673242023-01-22 Content Size-Dependent Alginate Microcapsule Formation Using Centrifugation to Eliminate Empty Microcapsules for On-Chip Imaging Cell Sorter Application Akimoto, Toshinosuke Yasuda, Kenji Micromachines (Basel) Article Alginate microcapsules are one of the attractive non-invasive platforms for handling individual cells and clusters, maintaining their isolation for further applications such as imaging cell sorter and single capsule qPCR. However, the conventional cell encapsulation techniques provide huge numbers of unnecessary empty homogeneous alginate microcapsules, which spend an excessive majority of the machine time on observations and analysis. Here, we developed a simple alginate cell encapsulation method to form content size-dependent alginate microcapsules to eliminate empty microcapsules using microcapillary centrifugation and filtration. Using this method, the formed calcium alginate microcapsules containing the HeLa cells were larger than 20m, and the other empty microcapsules were less than 3m under 4000 rpm centrifugation condition. We collected cell-containing alginate microcapsules by eliminating empty microcapsules from the microcapsule mixture with simple one-step filtration of a 20 m cell strainer. The electrical surface charge density and optical permeability of those cell-encapsulated alginate microcapsules were also evaluated. We found that the surface charge density of cell-encapsulated alginate microbeads is more than double that of cells, indicating that less voltage is required for electrical cell handling with thin alginate gel encapsulation of samples. The permeability of the alginate microcapsule was not improved by changing the reflective index of the medium buffer, such as adding alginate ester. However, the minimized thickness of the alginate gel envelope surrounding cells in the microcapsules did not degrade the detailed shapes of encapsulated cells. Those results confirmed the advantage of alginate encapsulation of cells with the centrifugation method as one of the desirable tools for imaging cell sorting applications. MDPI 2022-12-27 /pmc/articles/PMC9867324/ /pubmed/36677133 http://dx.doi.org/10.3390/mi14010072 Text en © 2022 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Akimoto, Toshinosuke
Yasuda, Kenji
Content Size-Dependent Alginate Microcapsule Formation Using Centrifugation to Eliminate Empty Microcapsules for On-Chip Imaging Cell Sorter Application
title Content Size-Dependent Alginate Microcapsule Formation Using Centrifugation to Eliminate Empty Microcapsules for On-Chip Imaging Cell Sorter Application
title_full Content Size-Dependent Alginate Microcapsule Formation Using Centrifugation to Eliminate Empty Microcapsules for On-Chip Imaging Cell Sorter Application
title_fullStr Content Size-Dependent Alginate Microcapsule Formation Using Centrifugation to Eliminate Empty Microcapsules for On-Chip Imaging Cell Sorter Application
title_full_unstemmed Content Size-Dependent Alginate Microcapsule Formation Using Centrifugation to Eliminate Empty Microcapsules for On-Chip Imaging Cell Sorter Application
title_short Content Size-Dependent Alginate Microcapsule Formation Using Centrifugation to Eliminate Empty Microcapsules for On-Chip Imaging Cell Sorter Application
title_sort content size-dependent alginate microcapsule formation using centrifugation to eliminate empty microcapsules for on-chip imaging cell sorter application
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9867324/
https://www.ncbi.nlm.nih.gov/pubmed/36677133
http://dx.doi.org/10.3390/mi14010072
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