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An integrated pipeline for high-throughput screening and profiling of spheroids using simple live image analysis of frame to frame variations
High-throughput imaging methods can be applied to relevant cell culture models, fostering their use in research and translational applications. Improvements in microscopy, computational capabilities and data analysis have enabled high-throughput, high-content approaches from endpoint 2D microscopy i...
Autores principales: | , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Academic Press
2021
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8165939/ https://www.ncbi.nlm.nih.gov/pubmed/32446959 http://dx.doi.org/10.1016/j.ymeth.2020.05.017 |
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author | Alsehli, Haneen Mosis, Fuad Thompson, Christopher Hamrud, Eva Wiseman, Erika Gentleman, Eileen Danovi, Davide |
author_facet | Alsehli, Haneen Mosis, Fuad Thompson, Christopher Hamrud, Eva Wiseman, Erika Gentleman, Eileen Danovi, Davide |
author_sort | Alsehli, Haneen |
collection | PubMed |
description | High-throughput imaging methods can be applied to relevant cell culture models, fostering their use in research and translational applications. Improvements in microscopy, computational capabilities and data analysis have enabled high-throughput, high-content approaches from endpoint 2D microscopy images. Nonetheless, trade-offs in acquisition, computation and storage between content and throughput remain, in particular when cells and cell structures are imaged in 3D. Moreover, live 3D phase contrast microscopy images are not often amenable to analysis because of the high level of background noise. Cultures of Human induced pluripotent stem cells (hiPSC) offer unprecedented scope to profile and screen conditions affecting cell fate decisions, self-organisation and early embryonic development. However, quantifying changes in the morphology or function of cell structures derived from hiPSCs over time presents significant challenges. Here, we report a novel method based on the analysis of live phase contrast microscopy images of hiPSC spheroids. We compare self-renewing versus differentiating media conditions, which give rise to spheroids with distinct morphologies; round versus branched, respectively. These cell structures are segmented from 2D projections and analysed based on frame-to-frame variations. Importantly, a tailored convolutional neural network is trained and applied to predict culture conditions from time-frame images. We compare our results with more classic and involved endpoint 3D confocal microscopy and propose that such approaches can complement spheroid-based assays developed for the purpose of screening and profiling. This workflow can be realistically implemented in laboratories using imaging-based high-throughput methods for regenerative medicine and drug discovery. |
format | Online Article Text |
id | pubmed-8165939 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2021 |
publisher | Academic Press |
record_format | MEDLINE/PubMed |
spelling | pubmed-81659392021-06-02 An integrated pipeline for high-throughput screening and profiling of spheroids using simple live image analysis of frame to frame variations Alsehli, Haneen Mosis, Fuad Thompson, Christopher Hamrud, Eva Wiseman, Erika Gentleman, Eileen Danovi, Davide Methods Article High-throughput imaging methods can be applied to relevant cell culture models, fostering their use in research and translational applications. Improvements in microscopy, computational capabilities and data analysis have enabled high-throughput, high-content approaches from endpoint 2D microscopy images. Nonetheless, trade-offs in acquisition, computation and storage between content and throughput remain, in particular when cells and cell structures are imaged in 3D. Moreover, live 3D phase contrast microscopy images are not often amenable to analysis because of the high level of background noise. Cultures of Human induced pluripotent stem cells (hiPSC) offer unprecedented scope to profile and screen conditions affecting cell fate decisions, self-organisation and early embryonic development. However, quantifying changes in the morphology or function of cell structures derived from hiPSCs over time presents significant challenges. Here, we report a novel method based on the analysis of live phase contrast microscopy images of hiPSC spheroids. We compare self-renewing versus differentiating media conditions, which give rise to spheroids with distinct morphologies; round versus branched, respectively. These cell structures are segmented from 2D projections and analysed based on frame-to-frame variations. Importantly, a tailored convolutional neural network is trained and applied to predict culture conditions from time-frame images. We compare our results with more classic and involved endpoint 3D confocal microscopy and propose that such approaches can complement spheroid-based assays developed for the purpose of screening and profiling. This workflow can be realistically implemented in laboratories using imaging-based high-throughput methods for regenerative medicine and drug discovery. Academic Press 2021-06 /pmc/articles/PMC8165939/ /pubmed/32446959 http://dx.doi.org/10.1016/j.ymeth.2020.05.017 Text en © 2020 The Authors https://creativecommons.org/licenses/by/4.0/This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/). |
spellingShingle | Article Alsehli, Haneen Mosis, Fuad Thompson, Christopher Hamrud, Eva Wiseman, Erika Gentleman, Eileen Danovi, Davide An integrated pipeline for high-throughput screening and profiling of spheroids using simple live image analysis of frame to frame variations |
title | An integrated pipeline for high-throughput screening and profiling of spheroids using simple live image analysis of frame to frame variations |
title_full | An integrated pipeline for high-throughput screening and profiling of spheroids using simple live image analysis of frame to frame variations |
title_fullStr | An integrated pipeline for high-throughput screening and profiling of spheroids using simple live image analysis of frame to frame variations |
title_full_unstemmed | An integrated pipeline for high-throughput screening and profiling of spheroids using simple live image analysis of frame to frame variations |
title_short | An integrated pipeline for high-throughput screening and profiling of spheroids using simple live image analysis of frame to frame variations |
title_sort | integrated pipeline for high-throughput screening and profiling of spheroids using simple live image analysis of frame to frame variations |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8165939/ https://www.ncbi.nlm.nih.gov/pubmed/32446959 http://dx.doi.org/10.1016/j.ymeth.2020.05.017 |
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