GCaMP is a genetically encoded calcium indicator that fluoresces in the presence of calcium ions, allowing for the visualization of neuronal activity.

How long does the imaging process take?

The imaging process can be conducted over a 24-hour period, capturing images every 15 to 30 minutes.

What type of microscope is used in this protocol?

A compound microscope equipped for wide field epifluorescence is used for calcium imaging.

Can this method be used for other organisms?

While this protocol is specific to C. elegans, similar techniques can be adapted for other model organisms expressing GCaMP.

What are the key steps in data analysis?

Data analysis involves assessing fluorescence intensity changes and using false color maps to enhance visibility of small changes.

Is this method suitable for beginners?

Yes, the protocol provides detailed steps that can be followed by researchers new to calcium imaging techniques.

Calcium Imaging: A Method to Visualize Neural Activity in Live C. elegans

JoVE Encyclopedia of Experiments

Biology

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- Place one transgenic worm expressing GCaMP-- a calcium sensing protein-- in an imaging chamber along with bacterial food for the worm. Then seal the dish with the lid and laboratory film to prevent evaporation. View it under a compound microscope equipped for wide field epifluorescence. Record time lapse images of the worms to track their behavior and capture fluorescence using imaging software.

Transgenic worms express a GCaMP sensor protein under the control of a promoter that drives GCaMP expression in a specific neuron. When that neuron is activated, it fires an action potential, which depolarizes the plasma membrane. Upon de-polarization, voltage-gated calcium channels open in the plasma membrane. This causes an influx of calcium ions into the cell, resulting in neuronal excitation.

GCaMP in the excited neuron binds the calcium ions. This causes GCaMP to fluoresce when the worms are imaged with a low intensity fluorescence excitation light. In the example protocol, we will use calcium imaging to visualize AVA interneuron activity in transgenic C. elegans in agarose microchambers.

- Calcium imaging is performed on a compound microscope equipped for wide field epifluorescence. To limit light exposure, use a transistor-transistor logic signal that triggers an LED to illuminate the sample at the same time the camera records a frame.

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Last updated: 27 June 2026

Calcium Imaging: A Method to Visualize Neural Activity in Live C. elegans

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