A Microfluidic System for Simultaneous Raman Spectroscopy, Patch-Clamp Electrophysiology, and Live-Cell Imaging to Study Key Cellular Events of Single Living Cells in Response to Acute Hypoxia.

Autor: Knoepp F; Excellence Cluster Cardio-Pulmonary Institute (CPI), University of Giessen and Marburg Lung Center (UGMLC), Member of the German Center for Lung Research (DZL), Justus-Liebig-University, Giessen, D-35392, Giessen, Germany., Wahl J; Department of Engineering Sciences and Mathematics, Luleå University of Technology, Luleå, SE-97187, Sweden., Andersson A; Department of Engineering Sciences and Mathematics, Luleå University of Technology, Luleå, SE-97187, Sweden., Kraut S; Excellence Cluster Cardio-Pulmonary Institute (CPI), University of Giessen and Marburg Lung Center (UGMLC), Member of the German Center for Lung Research (DZL), Justus-Liebig-University, Giessen, D-35392, Giessen, Germany., Sommer N; Excellence Cluster Cardio-Pulmonary Institute (CPI), University of Giessen and Marburg Lung Center (UGMLC), Member of the German Center for Lung Research (DZL), Justus-Liebig-University, Giessen, D-35392, Giessen, Germany., Weissmann N; Excellence Cluster Cardio-Pulmonary Institute (CPI), University of Giessen and Marburg Lung Center (UGMLC), Member of the German Center for Lung Research (DZL), Justus-Liebig-University, Giessen, D-35392, Giessen, Germany., Ramser K; Department of Engineering Sciences and Mathematics, Luleå University of Technology, Luleå, SE-97187, Sweden.
Jazyk: angličtina
Zdroj: Small methods [Small Methods] 2021 Oct; Vol. 5 (10), pp. e2100470. Date of Electronic Publication: 2021 Aug 21.
DOI: 10.1002/smtd.202100470
Abstrakt: The ability to sense changes in oxygen availability is fundamentally important for the survival of all aerobic organisms. However, cellular oxygen sensing mechanisms and pathologies remain incompletely understood and studies of acute oxygen sensing, in particular, have produced inconsistent results. Current methods cannot simultaneously measure the key cellular events in acute hypoxia (i.e., changes in redox state, electrophysiological properties, and mechanical responses) at controlled partial pressures of oxygen (pO 2 ). The lack of such a comprehensive method essentially contributes to the discrepancies in the field. A sealed microfluidic system that combines i) Raman spectroscopy, ii) patch-clamp electrophysiology, and iii) live-cell imaging under precisely controlled pO 2 have therefore been developed. Merging these modalities allows label-free and simultaneous observation of oxygen-dependent alterations in multiple cellular redox couples, membrane potential, and cellular contraction. This technique is adaptable to any cell type and allows in-depth insight into acute oxygen sensing processes underlying various physiologic and pathologic conditions.
(© 2021 The Authors. Small Methods published by Wiley-VCH GmbH.)
Databáze: MEDLINE
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