Click‐Chemistry‐Enabled Nanopipettes for the Capture and Dynamic Analysis of a Single Mitochondrion inside One Living Cell

Author:

Liu Kang1,Zhang Zheng1,Liu Rujia2,Li Jie P.3,Jiang Dechen1,Pan Rongrong1ORCID

Affiliation:

1. The State Key Lab of Analytical Chemistry for Life Science School of Chemistry and Chemical Engineering Chemistry and Biomedicine Innovation Center (ChemBIC) Nanjing University Nanjing Jiangsu 210093 China

2. School of Chemical Sciences University of Chinese Academy of Science Beijing 100190 China

3. The State Key Laboratory of Coordination Chemistry School of Chemistry and Chemical Engineering Chemistry and Biomedicine Innovation Center (ChemBIC) Nanjing University Nanjing Jiangsu 210093 China

Abstract

AbstractThe in‐depth study of single cells requires the dynamically molecular information in one particular nanometer‐sized organelle in a living cell, which is difficult to achieve using current methods. Due to high efficiency of click chemistry, a new nanoelectrode‐based pipette architecture with dibenzocyclooctyne at the tip is designed to realize fast conjugation with azide group‐containing triphenylphosphine, which targets mitochondrial membranes. The covalent binding of one mitochondrion at the tip of the nanopipette allows a small region of the membrane to be isolated on the Pt surface inside the nanopipette. Therefore, the release of reactive oxygen species (ROS) from the mitochondrion is monitored, which is not interfered by the species present in the cytosol. The dynamic tracking of ROS release from one mitochondrion reveals the distinctive “ROS‐induced ROS release” within the mitochondria. Further study of RSL3‐induced ferroptosis using nanopipettes provides direct evidence for supporting the noninvolvement of glutathione peroxidase 4 in the mitochondria during RSL3‐induced ROS generation, which has not previously been observed at the single‐mitochondrion level. Eventually, this established strategy should overcome the existing challenge of the dynamic measurement of one special organelle in the complicated intracellular environment, which opens a new direction for electroanalysis in subcellular analysis.

Funder

National Natural Science Foundation of China

China Postdoctoral Science Foundation

National Science Fund for Distinguished Young Scholars

Publisher

Wiley

Subject

General Medicine

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