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Robotic Intracellular Electrochemical Sensing for Adherent Cells

Nanopipette-based observation of intracellular biochemical processes is an important approach to revealing the intrinsic characteristics and heterogeneity of cells for better investigation of disease progression or early disease diagnosis. However, the manual operation needs a skilled operator and f...

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Detalles Bibliográficos
Autores principales: Hu, Weikang, Ma, Yanmei, Zhan, Zhen, Hussain, Danish, Hu, Chengzhi
Formato: Online Artículo Texto
Lenguaje:English
Publicado: AAAS 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9494721/
https://www.ncbi.nlm.nih.gov/pubmed/36285318
http://dx.doi.org/10.34133/2022/9763420
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author Hu, Weikang
Ma, Yanmei
Zhan, Zhen
Hussain, Danish
Hu, Chengzhi
author_facet Hu, Weikang
Ma, Yanmei
Zhan, Zhen
Hussain, Danish
Hu, Chengzhi
author_sort Hu, Weikang
collection PubMed
description Nanopipette-based observation of intracellular biochemical processes is an important approach to revealing the intrinsic characteristics and heterogeneity of cells for better investigation of disease progression or early disease diagnosis. However, the manual operation needs a skilled operator and faces problems such as low throughput and poor reproducibility. This paper proposes an automated nanopipette-based microoperation system for cell detection, three-dimensional nonovershoot positioning of the nanopipette tip in proximity to the cell of interest, cell approaching and proximity detection between nanopipette tip and cell surface, and cell penetration and detection of the intracellular reactive oxygen species (ROS). A robust focus algorithm based on the number of cell contours was proposed for adherent cells, which have sharp peaks while retaining unimodality. The automated detection of adherent cells was evaluated on human umbilical cord vein endothelial cells (HUVEC) and NIH/3T3 cells, which provided an average of 95.65% true-positive rate (TPR) and 7.59% false-positive rate (FPR) for in-plane cell detection. The three-dimensional nonovershoot tip positioning of the nanopipette was achieved by template matching and evaluated under the interference of cells. Ion current feedback was employed for the proximity detection between the nanopipette tip and cell surface. Finally, cell penetration and electrochemical detection of ROS were demonstrated on human breast cancer cells and zebrafish embryo cells. This work provides a systematic approach for automated intracellular sensing for adherent cells, laying a solid foundation for high-throughput detection, diagnosis, and classification of different forms of biochemical reactions within single cells.
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spelling pubmed-94947212022-10-24 Robotic Intracellular Electrochemical Sensing for Adherent Cells Hu, Weikang Ma, Yanmei Zhan, Zhen Hussain, Danish Hu, Chengzhi Cyborg Bionic Syst Research Article Nanopipette-based observation of intracellular biochemical processes is an important approach to revealing the intrinsic characteristics and heterogeneity of cells for better investigation of disease progression or early disease diagnosis. However, the manual operation needs a skilled operator and faces problems such as low throughput and poor reproducibility. This paper proposes an automated nanopipette-based microoperation system for cell detection, three-dimensional nonovershoot positioning of the nanopipette tip in proximity to the cell of interest, cell approaching and proximity detection between nanopipette tip and cell surface, and cell penetration and detection of the intracellular reactive oxygen species (ROS). A robust focus algorithm based on the number of cell contours was proposed for adherent cells, which have sharp peaks while retaining unimodality. The automated detection of adherent cells was evaluated on human umbilical cord vein endothelial cells (HUVEC) and NIH/3T3 cells, which provided an average of 95.65% true-positive rate (TPR) and 7.59% false-positive rate (FPR) for in-plane cell detection. The three-dimensional nonovershoot tip positioning of the nanopipette was achieved by template matching and evaluated under the interference of cells. Ion current feedback was employed for the proximity detection between the nanopipette tip and cell surface. Finally, cell penetration and electrochemical detection of ROS were demonstrated on human breast cancer cells and zebrafish embryo cells. This work provides a systematic approach for automated intracellular sensing for adherent cells, laying a solid foundation for high-throughput detection, diagnosis, and classification of different forms of biochemical reactions within single cells. AAAS 2022-09-02 /pmc/articles/PMC9494721/ /pubmed/36285318 http://dx.doi.org/10.34133/2022/9763420 Text en Copyright © 2022 Weikang Hu et al. https://creativecommons.org/licenses/by/4.0/Exclusive Licensee Beijing Institute of Technology Press. Distributed under a Creative Commons Attribution License (CC BY 4.0).
spellingShingle Research Article
Hu, Weikang
Ma, Yanmei
Zhan, Zhen
Hussain, Danish
Hu, Chengzhi
Robotic Intracellular Electrochemical Sensing for Adherent Cells
title Robotic Intracellular Electrochemical Sensing for Adherent Cells
title_full Robotic Intracellular Electrochemical Sensing for Adherent Cells
title_fullStr Robotic Intracellular Electrochemical Sensing for Adherent Cells
title_full_unstemmed Robotic Intracellular Electrochemical Sensing for Adherent Cells
title_short Robotic Intracellular Electrochemical Sensing for Adherent Cells
title_sort robotic intracellular electrochemical sensing for adherent cells
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9494721/
https://www.ncbi.nlm.nih.gov/pubmed/36285318
http://dx.doi.org/10.34133/2022/9763420
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