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the electrical properties of the tonoplast by Patch clamp (CAT#: STEM-PET-0062-WXH)
Introduction
A tonoplast is a cytoplasmic-membrane surrounding the vacuole within plant cells. The tonoplast is also known as the vacuolar membrane. The vacuole, found in the cytoplasm, prominent in the plant cells, is bound by a single membrane called tonoplast. Tonoplasts have special ion pumps and transporters that facilitate uphill transport of many ions and other materials, making their concentration higher in vacuole as compared to the cytoplasm.
Principle
The patch-clamp technique involves a glass micropipette forming a tight gigaohm seal with the cell membrane. The micropipette contains a wire bathed in an electrolytic solution to conduct ions. To measure single ion channels, a “patch” of membrane is pulled away from the cell after forming a gigaohm seal.
Applications
• Study of ionic currents in individual isolated living cells, tissue sections, or patches of cell membrane.
• Study of excitable cells such as neurons, cardiomyocytes, muscle fibers, and pancreatic beta cells.
• Study of ion channels.
• Study of excitable cells such as neurons, cardiomyocytes, muscle fibers, and pancreatic beta cells.
• Study of ion channels.
Procedure
1. Fabrication of glass electrodes
2. Measuring glass electrode resistance and compensating offset potential
3. Glass electrode contact to cell membrane and obtain a GΩ seal
4. Acquire and analyse recordings using the appropriate software.
2. Measuring glass electrode resistance and compensating offset potential
3. Glass electrode contact to cell membrane and obtain a GΩ seal
4. Acquire and analyse recordings using the appropriate software.
Materials
Patch clamp system
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