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Cultivation and Imaging of S. latissima Embryo Monolayered Cell Sheets Inside Microfluidic Devices
The culturing and investigation of individual marine specimens in lab environments is crucial to further our understanding of this highly complex ecosystem. However, the obtained results and their relevance are often limited by a lack of suitable experimental setups enabling controlled specimen grow...
Autores principales: | , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
MDPI
2022
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9687954/ https://www.ncbi.nlm.nih.gov/pubmed/36421119 http://dx.doi.org/10.3390/bioengineering9110718 |
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author | Clerc, Thomas Boscq, Samuel Attia, Rafaele Kaminski Schierle, Gabriele S. Charrier, Bénédicte Läubli, Nino F. |
author_facet | Clerc, Thomas Boscq, Samuel Attia, Rafaele Kaminski Schierle, Gabriele S. Charrier, Bénédicte Läubli, Nino F. |
author_sort | Clerc, Thomas |
collection | PubMed |
description | The culturing and investigation of individual marine specimens in lab environments is crucial to further our understanding of this highly complex ecosystem. However, the obtained results and their relevance are often limited by a lack of suitable experimental setups enabling controlled specimen growth in a natural environment while allowing for precise monitoring and in-depth observations. In this work, we explore the viability of a microfluidic device for the investigation of the growth of the alga Saccharina latissima to enable high-resolution imaging by confining the samples, which usually grow in 3D, to a single 2D plane. We evaluate the specimen’s health based on various factors such as its growth rate, cell shape, and major developmental steps with regard to the device’s operating parameters and flow conditions before demonstrating its compatibility with state-of-the-art microscopy imaging technologies such as the skeletonisation of the specimen through calcofluor white-based vital staining of its cell contours as well as the immunolocalisation of the specimen’s cell wall. Furthermore, by making use of the on-chip characterisation capabilities, we investigate the influence of altered environmental illuminations on the embryonic development using blue and red light. Finally, live tracking of fluorescent microspheres deposited on the surface of the embryo permits the quantitative characterisation of growth at various locations of the organism. |
format | Online Article Text |
id | pubmed-9687954 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2022 |
publisher | MDPI |
record_format | MEDLINE/PubMed |
spelling | pubmed-96879542022-11-25 Cultivation and Imaging of S. latissima Embryo Monolayered Cell Sheets Inside Microfluidic Devices Clerc, Thomas Boscq, Samuel Attia, Rafaele Kaminski Schierle, Gabriele S. Charrier, Bénédicte Läubli, Nino F. Bioengineering (Basel) Article The culturing and investigation of individual marine specimens in lab environments is crucial to further our understanding of this highly complex ecosystem. However, the obtained results and their relevance are often limited by a lack of suitable experimental setups enabling controlled specimen growth in a natural environment while allowing for precise monitoring and in-depth observations. In this work, we explore the viability of a microfluidic device for the investigation of the growth of the alga Saccharina latissima to enable high-resolution imaging by confining the samples, which usually grow in 3D, to a single 2D plane. We evaluate the specimen’s health based on various factors such as its growth rate, cell shape, and major developmental steps with regard to the device’s operating parameters and flow conditions before demonstrating its compatibility with state-of-the-art microscopy imaging technologies such as the skeletonisation of the specimen through calcofluor white-based vital staining of its cell contours as well as the immunolocalisation of the specimen’s cell wall. Furthermore, by making use of the on-chip characterisation capabilities, we investigate the influence of altered environmental illuminations on the embryonic development using blue and red light. Finally, live tracking of fluorescent microspheres deposited on the surface of the embryo permits the quantitative characterisation of growth at various locations of the organism. MDPI 2022-11-21 /pmc/articles/PMC9687954/ /pubmed/36421119 http://dx.doi.org/10.3390/bioengineering9110718 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 Clerc, Thomas Boscq, Samuel Attia, Rafaele Kaminski Schierle, Gabriele S. Charrier, Bénédicte Läubli, Nino F. Cultivation and Imaging of S. latissima Embryo Monolayered Cell Sheets Inside Microfluidic Devices |
title | Cultivation and Imaging of S. latissima Embryo Monolayered Cell Sheets Inside Microfluidic Devices |
title_full | Cultivation and Imaging of S. latissima Embryo Monolayered Cell Sheets Inside Microfluidic Devices |
title_fullStr | Cultivation and Imaging of S. latissima Embryo Monolayered Cell Sheets Inside Microfluidic Devices |
title_full_unstemmed | Cultivation and Imaging of S. latissima Embryo Monolayered Cell Sheets Inside Microfluidic Devices |
title_short | Cultivation and Imaging of S. latissima Embryo Monolayered Cell Sheets Inside Microfluidic Devices |
title_sort | cultivation and imaging of s. latissima embryo monolayered cell sheets inside microfluidic devices |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9687954/ https://www.ncbi.nlm.nih.gov/pubmed/36421119 http://dx.doi.org/10.3390/bioengineering9110718 |
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