Abstract
We developed a highly sensitive oxygen consumption scanning microscopy system using platinized platinum disc microelectrodes. The system is capable of reliably detecting single-cell respiration, responding to classical regulators of mitochondrial oxygen consumption activity as expected. Comparisons with commercial multi-cell oxygen detection systems show that the system has comparable errors (if not smaller), with the advantage of being able to monitor inter and intra-cell heterogeneity in oxygen consumption characteristics. Our results uncover heterogeneous oxygen consumption characteristics between cells and within the same cell´s microenvironments. Single Cell Oxygen Mapping (SCOM) is thus capable of reliably studying mitochondrial oxygen consumption characteristics and heterogeneity at a single-cell level.
Highlights
We believe the area would greatly benefit from the development of single cell oxygen consumption techniques
Typical O2 electrochemical detection responses of the constructed sensor can be seen in Fig. 1, which shows cyclic voltammograms recorded in the absence of O2 (Fig. 1A, black line, i) air-saturated solution (Fig. 1A, red curve, ii) and O2-saturated phosphate buffered saline solution (Fig. 1A, blue curve, iii)
A steady-state situation is achieved in curves ii and iii, which correspond to the electrochemical process involving O2 reduction
Summary
We believe the area would greatly benefit from the development of single cell oxygen consumption techniques. Different techniques have been used to acquire topographical information with high spatial resolution, including atomic force microscopy (AFM), scanning electron microscopy (SEM) and scanning electrochemical microscopy (SECM), which is highly valuable in measurements of local electrochemical activity at interfaces[13,14,15,16]. We present an effective and simple approach to evaluate oxygen consumption in the microenvironment of an individual cell, using a platinized platinum disc microelectrode as a tip in SECM configuration. The use of a platinized platinum microelectrode as a tip in a SECM configuration for mapping the oxygen concentration above a single-cell uncovers rich topographical heterogeneity in oxygen uptake characteristics within individual cells in culture, which may have important regulatory, physiological and pathological implications
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