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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Main Authors: Yuanjun Zhang, Zishen Yan, Xingyu Xia, Yuan Lin
Format: Article
Language:English
Published: MDPI AG 2020-08-01
Series:Micromachines
Subjects:
Online Access:https://www.mdpi.com/2072-666X/11/8/767
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spelling 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
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