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Enzyme-histochemical method for identification of lymphatic capillaries
1Department of Anatomy, Medical College of Oita, Japan.
Lymphology
|September 1, 1991
Summary
This study distinguishes lymphatic and blood capillaries using enzyme histochemistry. 5'-nucleotidase (5'-Nase) identifies lymphatic capillaries, while alkaline phosphatase (ALPase) identifies blood capillaries in monkey intestines.
Area of Science:
- Histochemistry
- Cell Biology
- Microscopy
Background:
- Distinguishing between lymphatic and blood capillaries is crucial for understanding microcirculation.
- Traditional methods may lack specificity or resolution.
Purpose of the Study:
- To develop and validate a double staining method for differentiating lymphatic and blood capillaries.
- To characterize the enzyme activity and localization in these microvessels.
Main Methods:
- Utilized 5 acutenucleotidase (5 acutenase) and alkaline phosphatase (ALPase) double staining on glycol methacrylate sections.
- Employed L-tetramisole to inhibit nonspecific ALPase, ensuring 5 acutenase specificity.
- Applied light and transmission electron microscopy for visualization and localization of enzyme activity.
Main Results:
- Achieved distinct visualization of 5 acutenase activity in lymphatic capillaries and ALPase activity in blood capillaries.
- Demonstrated intense 5 acutenase activity on the outer plasma membrane of lymphatic endothelial cells.
- Confirmed the specificity of the enzyme reactions for capillary differentiation.
Conclusions:
- The 5 acutenase-ALPase double staining method reliably distinguishes lymphatic from blood capillaries.
- This technique provides detailed insights into the enzyme activity and ultrastructural localization within capillary walls.
- The findings enhance our understanding of microvascular structures and their functions.