Determination of single-cell oxygen consumption with impedance feedback for control of sample-probe separation.
The ability to measure chemical gradients surrounding single cells provides novel insights into several areas of cell dynamics--particularly metabolism. Detection of metabolic oxygen consumption can be achieved from a single mammalian cell using a modulated amperometric sensor in a self-referencing mode. To date, however, apart from visual cues, we do not have a reliable and cell-compatible method for determining and stabilizing the position of such probes. In this paper, we report on having successfully measured the increase in the uncompensated resistance of an electrochemical cell upon approach to single, living, biological cells, while simultaneously measuring the metabolic oxygen consumption. This was accomplished by applying an ac and a dc excitation signal to the electrode. The applied ac waveform was a 100-kHz sine wave with an amplitude of 10 mV rms, while the dc voltage applied was -600 mV. The two signals were shown not to interfere with one another. Furthermore, it is shown that the sample-probe distance can be measured for approach to single cells on the order of 10-15-microm diameter and 5-microm height, with 100-nm resolution.
Osbourn
Damon M
DM
Program in Molecular Physiology, Marine Biological Laboratory, BioCurrents Research Center, Woods Hole, Massachusetts 02543, USA.
Sanger
Richard H
RH
Smith
Peter J S
PJ
D000818:Animals; D002460:Cell Line; D003584:Cytological Techniques / Q000295:instrumentation / Q000379:methods*; D017097:Electric Impedance; D007340:Insulinoma; D010101:Oxygen Consumption / Q000502:physiology*; D051381:Rats