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Cell sorting in a Petri dish controlled by computer vision

Fluorescence-activated cell sorting (FACS) applying flow cytometry to separate cells on a molecular basis is a widespread method. We demonstrate that both fluorescent and unlabeled live cells in a Petri dish observed with a microscope can be automatically recognized by computer vision and picked up...

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Autores principales: Környei, Z., Beke, S., Mihálffy, T., Jelitai, M., Kovács, K. J., Szabó, Z., Szabó, B.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Nature Publishing Group 2013
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3548191/
https://www.ncbi.nlm.nih.gov/pubmed/23336070
http://dx.doi.org/10.1038/srep01088
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author Környei, Z.
Beke, S.
Mihálffy, T.
Jelitai, M.
Kovács, K. J.
Szabó, Z.
Szabó, B.
author_facet Környei, Z.
Beke, S.
Mihálffy, T.
Jelitai, M.
Kovács, K. J.
Szabó, Z.
Szabó, B.
author_sort Környei, Z.
collection PubMed
description Fluorescence-activated cell sorting (FACS) applying flow cytometry to separate cells on a molecular basis is a widespread method. We demonstrate that both fluorescent and unlabeled live cells in a Petri dish observed with a microscope can be automatically recognized by computer vision and picked up by a computer-controlled micropipette. This method can be routinely applied as a FACS down to the single cell level with a very high selectivity. Sorting resolution, i.e., the minimum distance between two cells from which one could be selectively removed was 50–70 micrometers. Survival rate with a low number of 3T3 mouse fibroblasts and NE-4C neuroectodermal mouse stem cells was 66±12% and 88±16%, respectively. Purity of sorted cultures and rate of survival using NE-4C/NE-GFP-4C co-cultures were 95±2% and 62±7%, respectively. Hydrodynamic simulations confirmed the experimental sorting efficiency and a cell damage risk similar to that of normal FACS.
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spelling pubmed-35481912013-01-18 Cell sorting in a Petri dish controlled by computer vision Környei, Z. Beke, S. Mihálffy, T. Jelitai, M. Kovács, K. J. Szabó, Z. Szabó, B. Sci Rep Article Fluorescence-activated cell sorting (FACS) applying flow cytometry to separate cells on a molecular basis is a widespread method. We demonstrate that both fluorescent and unlabeled live cells in a Petri dish observed with a microscope can be automatically recognized by computer vision and picked up by a computer-controlled micropipette. This method can be routinely applied as a FACS down to the single cell level with a very high selectivity. Sorting resolution, i.e., the minimum distance between two cells from which one could be selectively removed was 50–70 micrometers. Survival rate with a low number of 3T3 mouse fibroblasts and NE-4C neuroectodermal mouse stem cells was 66±12% and 88±16%, respectively. Purity of sorted cultures and rate of survival using NE-4C/NE-GFP-4C co-cultures were 95±2% and 62±7%, respectively. Hydrodynamic simulations confirmed the experimental sorting efficiency and a cell damage risk similar to that of normal FACS. Nature Publishing Group 2013-01-18 /pmc/articles/PMC3548191/ /pubmed/23336070 http://dx.doi.org/10.1038/srep01088 Text en Copyright © 2013, Macmillan Publishers Limited. All rights reserved http://creativecommons.org/licenses/by-nc-nd/3.0/ This work is licensed under a Creative Commons Attribution-NonCommercial-NoDerivs 3.0 Unported License. To view a copy of this license, visit http://creativecommons.org/licenses/by-nc-nd/3.0/
spellingShingle Article
Környei, Z.
Beke, S.
Mihálffy, T.
Jelitai, M.
Kovács, K. J.
Szabó, Z.
Szabó, B.
Cell sorting in a Petri dish controlled by computer vision
title Cell sorting in a Petri dish controlled by computer vision
title_full Cell sorting in a Petri dish controlled by computer vision
title_fullStr Cell sorting in a Petri dish controlled by computer vision
title_full_unstemmed Cell sorting in a Petri dish controlled by computer vision
title_short Cell sorting in a Petri dish controlled by computer vision
title_sort cell sorting in a petri dish controlled by computer vision
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3548191/
https://www.ncbi.nlm.nih.gov/pubmed/23336070
http://dx.doi.org/10.1038/srep01088
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