简介：
Overview
This article presents protocols for using the genetically-encoded calcium indicator (GECI) CatchER + to monitor rapid calcium transients in HEK293 and C2C12 cells. The methods include real-time fluorescence microscopy and in situ K d measurement for calibration.
Key Study Components
Area of Science
- Neuroscience
- Cell Biology
- Fluorescence Microscopy
Background
- Calcium signaling is crucial for various cellular processes.
- Monitoring calcium transients can provide insights into cellular health.
- CatchER + is a novel tool for studying calcium dynamics.
- Real-time imaging allows for the observation of rapid changes in calcium levels.
Purpose of Study
- To demonstrate protocols for cell culture and transfection with CatchER +.
- To measure ER/SR calcium transients in HEK293 and C2C12 cells.
- To explore the implications of calcium signaling in disease.
Methods Used
- Cell culture of HEK293 and C2C12 cells.
- Transfection with the CatchER + biosensor.
- Real-time fluorescence microscopy for imaging calcium transients.
- Calibration of the biosensor using in situ K d measurement.
Main Results
- Successful transfection of cells with CatchER +.
- Real-time monitoring of calcium transients in the ER/SR.
- Demonstration of rapid kinetics in calcium signaling.
- Potential applications in diagnosing calcium-related diseases.
Conclusions
- CatchER + is effective for monitoring calcium dynamics in live cells.
- This technique can enhance understanding of calcium signaling.
- Visual demonstrations are crucial for teaching complex protocols.
What is CatchER +?
CatchER + is a genetically-encoded calcium indicator used to monitor calcium transients in cells.
How does real-time fluorescence microscopy work?
It allows for the visualization of dynamic processes in live cells by detecting fluorescence emitted from the biosensor.
What types of cells were used in this study?
HEK293 and C2C12 cells were used for the experiments.
Why is monitoring calcium transients important?
It provides insights into cellular signaling and can help identify dysfunctions related to diseases.
What are the advantages of using CatchER +?
It allows for real-time monitoring of calcium dynamics with rapid kinetics, enhancing experimental accuracy.
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