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...
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MDPI AG
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doaj-42a116f15f9f4bb5a774c8ff9ee1fdd12020-11-25T03:07:23ZengMDPI AGMicromachines2072-666X2020-08-011176776710.3390/mi11080767A Novel Electroporation System for Living Cell Staining and Membrane Dynamics InterrogationYuanjun Zhang0Zishen Yan1Xingyu Xia2Yuan Lin3HKU-Shenzhen Institute of Research and Innovation (HKU-SIRI), Guangdong 518057, ChinaHKU-Shenzhen Institute of Research and Innovation (HKU-SIRI), Guangdong 518057, ChinaHKU-Shenzhen Institute of Research and Innovation (HKU-SIRI), Guangdong 518057, ChinaDepartment of Mechanical Engineering, The University of Hong Kong, Hong Kong SAR, ChinaA 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.https://www.mdpi.com/2072-666X/11/8/767electroporationmembrane resealingelectrotransfectionlive cell stainingcell adhesion |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Yuanjun Zhang Zishen Yan Xingyu Xia Yuan Lin |
spellingShingle |
Yuanjun Zhang Zishen Yan Xingyu Xia Yuan Lin A Novel Electroporation System for Living Cell Staining and Membrane Dynamics Interrogation Micromachines electroporation membrane resealing electrotransfection live cell staining cell adhesion |
author_facet |
Yuanjun Zhang Zishen Yan Xingyu Xia Yuan Lin |
author_sort |
Yuanjun Zhang |
title |
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_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_sort |
novel electroporation system for living cell staining and membrane dynamics interrogation |
publisher |
MDPI AG |
series |
Micromachines |
issn |
2072-666X |
publishDate |
2020-08-01 |
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. |
topic |
electroporation membrane resealing electrotransfection live cell staining cell adhesion |
url |
https://www.mdpi.com/2072-666X/11/8/767 |
work_keys_str_mv |
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