全文：
Overview
This article presents a protocol for measuring the induced membrane voltage (ΔΦ) in cells using the potentiometric dye di-8-ANEPPS. The induced voltage occurs when a cell is placed in an electric field, leading to charge redistribution across the membrane.
Key Study Components
Area of Science
- Electrophysiology
- Cell Biology
- Membrane Dynamics
Background
- Cells are conductive structures enclosed by an insulating membrane.
- The resting membrane voltage is influenced by ionic concentration imbalances.
- Induced membrane voltage can be calculated or measured experimentally.
- Di-8-ANEPPS is a dye used for noninvasive measurement of membrane voltage changes.
Purpose of Study
- To demonstrate a method for measuring induced membrane voltage in cells.
- To illustrate the use of di-8-ANEPPS in fluorescence microscopy.
- To provide a detailed protocol for researchers to follow.
Methods Used
- Preparation of cell cultures in specific media.
- Loading cells with di-8-ANEPPS dye.
- Application of electric pulses to induce membrane voltage.
- Fluorescence imaging to measure changes in membrane voltage.
Main Results
- The protocol allows for the measurement of induced membrane voltage in various cell shapes.
- Fluorescence intensity changes correlate with induced voltage levels.
- Data processing enables accurate determination of membrane voltage.
- Results align with theoretical predictions for spherical cells.
Conclusions
- The method provides a reliable approach for studying membrane voltage dynamics.
- Di-8-ANEPPS is effective for noninvasive measurements in live cells.
- Further calibration may enhance measurement accuracy for different setups.
What is induced membrane voltage?
Induced membrane voltage (ΔΦ) is the voltage change across a cell membrane when exposed to an external electric field.
How is di-8-ANEPPS used in this study?
Di-8-ANEPPS is a potentiometric dye that allows for the noninvasive measurement of changes in membrane voltage.
What type of cells were used in the experiments?
Chinese hamster ovary (CHO) cells were used for the experiments.
What is the significance of measuring membrane voltage?
Measuring membrane voltage is crucial for understanding cellular excitability and signaling mechanisms.
What equipment is needed for this protocol?
A fluorescence microscope, a pulse generator, and specific culture media are required for the protocol.
How can the results be validated?
Results can be validated by comparing them with theoretical predictions and using calibration curves for accuracy.
繁體中文
