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Generating kinetic environments to study dynamic cellular processes in single cells

Cells of any organism are consistently exposed to changes over time in their environment. The kinetics by which these changes occur are critical for the cellular response and fate decision. It is therefore important to control the temporal changes of extracellular stimuli precisely to understand bio...

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Autores principales: Thiemicke, Alexander, Jashnsaz, Hossein, Li, Guoliang, Neuert, Gregor
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Nature Publishing Group UK 2019
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6625993/
https://www.ncbi.nlm.nih.gov/pubmed/31300695
http://dx.doi.org/10.1038/s41598-019-46438-8
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author Thiemicke, Alexander
Jashnsaz, Hossein
Li, Guoliang
Neuert, Gregor
author_facet Thiemicke, Alexander
Jashnsaz, Hossein
Li, Guoliang
Neuert, Gregor
author_sort Thiemicke, Alexander
collection PubMed
description Cells of any organism are consistently exposed to changes over time in their environment. The kinetics by which these changes occur are critical for the cellular response and fate decision. It is therefore important to control the temporal changes of extracellular stimuli precisely to understand biological mechanisms in a quantitative manner. Most current cell culture and biochemical studies focus on instant changes in the environment and therefore neglect the importance of kinetic environments. To address these shortcomings, we developed two experimental methodologies to precisely control the environment of single cells. These methodologies are compatible with standard biochemistry, molecular, cell and quantitative biology assays. We demonstrate applicability by obtaining time series and time point measurements in both live and fixed cells. We demonstrate the feasibility of the methodology in yeast and mammalian cell culture in combination with widely used assays such as flow cytometry, time-lapse microscopy and single-molecule RNA Fluorescent in-situ Hybridization (smFISH). Our experimental methodologies are easy to implement in most laboratory settings and allows the study of kinetic environments in a wide range of assays and different cell culture conditions.
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spelling pubmed-66259932019-07-21 Generating kinetic environments to study dynamic cellular processes in single cells Thiemicke, Alexander Jashnsaz, Hossein Li, Guoliang Neuert, Gregor Sci Rep Article Cells of any organism are consistently exposed to changes over time in their environment. The kinetics by which these changes occur are critical for the cellular response and fate decision. It is therefore important to control the temporal changes of extracellular stimuli precisely to understand biological mechanisms in a quantitative manner. Most current cell culture and biochemical studies focus on instant changes in the environment and therefore neglect the importance of kinetic environments. To address these shortcomings, we developed two experimental methodologies to precisely control the environment of single cells. These methodologies are compatible with standard biochemistry, molecular, cell and quantitative biology assays. We demonstrate applicability by obtaining time series and time point measurements in both live and fixed cells. We demonstrate the feasibility of the methodology in yeast and mammalian cell culture in combination with widely used assays such as flow cytometry, time-lapse microscopy and single-molecule RNA Fluorescent in-situ Hybridization (smFISH). Our experimental methodologies are easy to implement in most laboratory settings and allows the study of kinetic environments in a wide range of assays and different cell culture conditions. Nature Publishing Group UK 2019-07-12 /pmc/articles/PMC6625993/ /pubmed/31300695 http://dx.doi.org/10.1038/s41598-019-46438-8 Text en © The Author(s) 2019 Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.
spellingShingle Article
Thiemicke, Alexander
Jashnsaz, Hossein
Li, Guoliang
Neuert, Gregor
Generating kinetic environments to study dynamic cellular processes in single cells
title Generating kinetic environments to study dynamic cellular processes in single cells
title_full Generating kinetic environments to study dynamic cellular processes in single cells
title_fullStr Generating kinetic environments to study dynamic cellular processes in single cells
title_full_unstemmed Generating kinetic environments to study dynamic cellular processes in single cells
title_short Generating kinetic environments to study dynamic cellular processes in single cells
title_sort generating kinetic environments to study dynamic cellular processes in single cells
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6625993/
https://www.ncbi.nlm.nih.gov/pubmed/31300695
http://dx.doi.org/10.1038/s41598-019-46438-8
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