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Oxygen microsensor and its application to single cells and mouse pancreatic islets
S K Jung1, W Gorski, C A Aspinwall
1Department of Chemistry, University of Florida, Gainesville 32611-7200, USA.
Analytical Chemistry
|September 18, 1999
Summary
A novel oxygen microsensor monitors single cells and pancreatic islets. This tool reveals oxygen dynamics and consumption, confirming adequate oxygen for insulin secretion in islets under culture conditions.
Area of Science:
- Biomedical Engineering
- Cell Biology
- Physiology
Background:
- Accurate monitoring of oxygen levels is crucial for understanding cellular metabolism and function.
- Existing oxygen sensors often lack the spatial resolution required for single-cell or micro-tissue analysis.
- Pancreatic islets' oxygenation is vital for insulin secretion, but dynamic changes are poorly understood.
Purpose of the Study:
- To develop and characterize a high-resolution oxygen microsensor for biological microenvironments.
- To investigate oxygen gradients and dynamics within mouse pancreatic islets.
- To measure oxygen consumption in single insulinoma cells and assess islet oxygenation under varying glucose conditions.
Main Methods:
- Fabrication of a sub-3-micron tip diameter oxygen microsensor using electrochemistry and glass micropipette technology.
- Characterization of sensor sensitivity, response time, and convective perturbation.
- In situ measurement of oxygen levels and gradients in single mouse pancreatic islets and insulinoma cells.
- Dynamic recording of oxygen levels in response to glucose concentration changes and observation of oscillatory patterns.
Main Results:
- The developed microsensor exhibited high sensitivity (< 1 s response time) and minimal convection perturbation.
- Oxygen levels within mouse islets were sufficient for insulin secretion under tissue culture conditions (67 +/- 1.6 mmHg).
- Glucose stimulation induced oxygen level decreases and oscillations (3.3 +/- 0.6 min periods) in islets, with faster oscillations (12.1 +/- 1.7 s) also detected.
- Single insulinoma cells demonstrated dynamic oxygen consumption with rapid oscillations.
Conclusions:
- The novel oxygen microsensor provides a powerful tool for dynamic, high-resolution oxygen monitoring in biological microenvironments.
- Islets possess adequate oxygen diffusion under standard culture conditions, supporting insulin secretion.
- Glucose metabolism in islets and single cells is associated with dynamic oxygen level fluctuations and consumption oscillations, offering insights into cellular energetics.