| RRC ID |
64536
|
| Author |
Subramanian Parimalam S, Abdelmoez MN, Tsuchida A, Sotta N, Tanaka M, Kuromori T, Fujiwara T, Hirai MY, Yokokawa R, Oguchi Y, Shintaku H.
|
| Title |
Targeted permeabilization of the cell wall and extraction of charged molecules from single cells in intact plant clusters using a focused electric field.
|
| Journal |
Analyst
|
| 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.
|
| Volume |
146(5)
|
| Pages |
1604-1611
|
| Published |
2021-3-7
|
| DOI |
10.1039/d0an02163f
|
| PMID |
33624642
|
| MeSH |
Cell Wall
Isotachophoresis*
Microfluidics
Oligonucleotide Array Sequence Analysis
Plants
|
| IF |
3.978
|
| Resource |
| Arabidopsis / Cultured plant cells, genes |
rpc00105 |
