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Intracellular ion imaging using fluorescent dyes: artefacts and limits to resolution
R A Silver1, M Whitaker, S R Bolsover
1Department of Physiology, University College London, UK.
Pflugers Archiv : European Journal of Physiology
|April 1, 1992
Summary
This study explores the challenges and potential artifacts in cellular ion concentration imaging using fluorescent ratio indicators. It details methods to improve imaging time resolution and minimize errors for more accurate cellular analysis.
Area of Science:
- Cellular Biology
- Biophysics
- Microscopy Techniques
Background:
- Fluorescent ratio indicators enable cellular ion concentration imaging.
- Ion imaging is complex and susceptible to artifacts.
- Understanding limitations is crucial for accurate data.
Purpose of the Study:
- Investigate the limits and pitfalls of ion imaging techniques.
- Enhance the time resolution of existing ion imaging systems.
- Identify and provide solutions for common imaging artifacts.
Main Methods:
- Analysis of spatial resolution across different cell geometries.
- Development of a single-excitation wavelength technique for improved time resolution.
- Demonstration of artifacts from hardware, image processing, and optics.
Main Results:
- Spatial resolution is dependent on cell geometry.
- A novel technique enhances time resolution using a single excitation wavelength.
- Common artifacts were identified and exemplified.
Conclusions:
- Accurate cellular ion imaging requires careful consideration of technique limitations.
- Methods for artifact recognition and minimization are essential for reliable results.
- Improved techniques can enhance the utility of ion imaging in biological research.
