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Issue 5, 2021
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Targeted permeabilization of the cell wall and extraction of charged molecules from single cells in intact plant clusters using a focused electric field

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Abstract

The extraction of cellular contents from plant cells covered with cell walls remains a challenge, as it is physically hindered by the cell wall. We present a new microfluidic approach that leverages an intense pulsed electric field for permeabilizing the cell wall and a focused DC electric field for extracting the cellular contents selectively from a few targeted cells in a cluster of intact plant cells. We coupled the approach with on-chip fluorescence quantification of extracted molecules leveraging isotachophoresis as well as off-chip reverse transcription-quantitative polymerase chain reaction detecting extracted mRNA molecules. Our approach offers a workflow of about 5 min, isolating a cluster of intact plant cells, permeabilizing the cell wall, selectively extracting cytosolic molecules from a few targeted cells in the cluster, and outputting them to off-chip analyses without any enzymatic reactions. We anticipate that this approach will create a new opportunity to explore plant biology through less biased data realized by the rapid extraction of molecules from intact plant clusters.

Graphical abstract: Targeted permeabilization of the cell wall and extraction of charged molecules from single cells in intact plant clusters using a focused electric field

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Article information


Submitted
01 Nov 2020
Accepted
08 Feb 2021
First published
11 Feb 2021

Analyst, 2021,146, 1604-1611
Article type
Paper

Targeted permeabilization of the cell wall and extraction of charged molecules from single cells in intact plant clusters using a focused electric field

S. Subramanian Parimalam, M. N. Abdelmoez, A. Tsuchida, N. Sotta, M. Tanaka, T. Kuromori, T. Fujiwara, M. Y. Hirai, R. Yokokawa, Y. Oguchi and H. Shintaku, Analyst, 2021, 146, 1604
DOI: 10.1039/D0AN02163F

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