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Quantitative laser scanning confocal microscopy on single capillaries: permeability measurement
R H Adamson1, J F Lenz, F E Curry
1Department of Human Physiology, School of Medicine, University of California 95616, USA.
Microcirculation (New York, N.Y. : 1994)
|December 1, 1994
Summary
Solute loss to superfusate does not significantly affect microvessel permeability measurements in frog mesentery. This study utilized laser-scanning confocal microscopy to accurately assess permeability coefficients.
Area of Science:
- Physiology
- Microcirculation Research
- Confocal Microscopy Applications
Background:
- Accurate measurement of microvessel permeability is crucial for understanding physiological and pathological processes.
- Previous methods may have been confounded by solute loss to the superfusate.
- Laser-scanning confocal microscopy offers a novel approach for high-resolution microvessel analysis.
Purpose of the Study:
- To test if solute loss to the superfusate affects microvessel permeability measurements in frog mesentery.
- To determine if permeability coefficients are underestimated due to superfusate solute loss.
- To validate the use of laser-scanning confocal microscopy for microvessel permeability studies.
Main Methods:
- Microvessel permeability to sodium fluorescein was measured using laser-scanning confocal microscopy in frog mesentery.
- Measurements were conducted with Ringer's solution superfusion and subsequently with mineral oil superfusion to assess solute retention.
- Image analysis involved corrections for fluorescence attenuation and light collection nonuniformity.
Main Results:
- Microvessel permeability to fluorescein was similar when superfused with Ringer's solution (5.0 ± 2.6 x 10⁻⁵ cm/s) compared to mineral oil (4.5 ± 1.9 x 10⁻⁵ cm/s).
- No significant difference in permeability was observed between the two superfusion conditions.
- These findings indicate that sodium fluorescein loss through the mesothelium did not significantly alter permeability under the tested conditions.
Conclusions:
- Solute loss to the superfusate does not significantly impact microvessel permeability measurements in frog mesentery under these experimental conditions.
- The employed laser-scanning confocal microscopy techniques provide a robust method for detailed analysis of solute concentration gradients around microvessels.
- This study validates the accuracy of permeability measurements using this advanced imaging technique.