Cargando…
A Novel Electroporation System for Living Cell Staining and Membrane Dynamics Interrogation
A novel electroporation system was developed to introduce transient membrane pores to cells in a spatially and temporally controlled manner, allowing us to achieve fast electrotransfection and live cell staining as well as to systematically interrogate the dynamics of the cell membrane. Specifically...
Autores principales: | , , , |
---|---|
Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
MDPI
2020
|
Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7466103/ https://www.ncbi.nlm.nih.gov/pubmed/32796554 http://dx.doi.org/10.3390/mi11080767 |
_version_ | 1783577735525826560 |
---|---|
author | Zhang, Yuanjun Yan, Zishen Xia, Xingyu Lin, Yuan |
author_facet | Zhang, Yuanjun Yan, Zishen Xia, Xingyu Lin, Yuan |
author_sort | Zhang, Yuanjun |
collection | PubMed |
description | A novel electroporation system was developed to introduce transient membrane pores to cells in a spatially and temporally controlled manner, allowing us to achieve fast electrotransfection and live cell staining as well as to systematically interrogate the dynamics of the cell membrane. Specifically, using this platform, we showed that both reversible and irreversible electroporation could be induced in the cell population, with nano-sized membrane pores in the former case being able to self-reseal in ~10 min. In addition, green fluorescent protein(GFP)-vinculin plasmid and 543 phalloidin have been delivered successively into fibroblast cells, which enables us to monitor the distinct roles of vinculin and F-actin in cell adhesion and migration as well as their possible interplay during these processes. Compared to conventional bulk electroporation and staining methods, the new system offers advantages such as low-voltage operation, cellular level manipulation and testing, fast and adjustable transfection/staining and real-time monitoring; the new system therefore could be useful in different biophysical studies in the future. |
format | Online Article Text |
id | pubmed-7466103 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2020 |
publisher | MDPI |
record_format | MEDLINE/PubMed |
spelling | pubmed-74661032020-09-14 A Novel Electroporation System for Living Cell Staining and Membrane Dynamics Interrogation Zhang, Yuanjun Yan, Zishen Xia, Xingyu Lin, Yuan Micromachines (Basel) Article A novel electroporation system was developed to introduce transient membrane pores to cells in a spatially and temporally controlled manner, allowing us to achieve fast electrotransfection and live cell staining as well as to systematically interrogate the dynamics of the cell membrane. Specifically, using this platform, we showed that both reversible and irreversible electroporation could be induced in the cell population, with nano-sized membrane pores in the former case being able to self-reseal in ~10 min. In addition, green fluorescent protein(GFP)-vinculin plasmid and 543 phalloidin have been delivered successively into fibroblast cells, which enables us to monitor the distinct roles of vinculin and F-actin in cell adhesion and migration as well as their possible interplay during these processes. Compared to conventional bulk electroporation and staining methods, the new system offers advantages such as low-voltage operation, cellular level manipulation and testing, fast and adjustable transfection/staining and real-time monitoring; the new system therefore could be useful in different biophysical studies in the future. MDPI 2020-08-11 /pmc/articles/PMC7466103/ /pubmed/32796554 http://dx.doi.org/10.3390/mi11080767 Text en © 2020 by the authors. 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 (http://creativecommons.org/licenses/by/4.0/). |
spellingShingle | Article Zhang, Yuanjun Yan, Zishen Xia, Xingyu Lin, Yuan A Novel Electroporation System for Living Cell Staining and Membrane Dynamics Interrogation |
title | A Novel Electroporation System for Living Cell Staining and Membrane Dynamics Interrogation |
title_full | A Novel Electroporation System for Living Cell Staining and Membrane Dynamics Interrogation |
title_fullStr | A Novel Electroporation System for Living Cell Staining and Membrane Dynamics Interrogation |
title_full_unstemmed | A Novel Electroporation System for Living Cell Staining and Membrane Dynamics Interrogation |
title_short | A Novel Electroporation System for Living Cell Staining and Membrane Dynamics Interrogation |
title_sort | novel electroporation system for living cell staining and membrane dynamics interrogation |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7466103/ https://www.ncbi.nlm.nih.gov/pubmed/32796554 http://dx.doi.org/10.3390/mi11080767 |
work_keys_str_mv | AT zhangyuanjun anovelelectroporationsystemforlivingcellstainingandmembranedynamicsinterrogation AT yanzishen anovelelectroporationsystemforlivingcellstainingandmembranedynamicsinterrogation AT xiaxingyu anovelelectroporationsystemforlivingcellstainingandmembranedynamicsinterrogation AT linyuan anovelelectroporationsystemforlivingcellstainingandmembranedynamicsinterrogation AT zhangyuanjun novelelectroporationsystemforlivingcellstainingandmembranedynamicsinterrogation AT yanzishen novelelectroporationsystemforlivingcellstainingandmembranedynamicsinterrogation AT xiaxingyu novelelectroporationsystemforlivingcellstainingandmembranedynamicsinterrogation AT linyuan novelelectroporationsystemforlivingcellstainingandmembranedynamicsinterrogation |