Simultaneous imaging of morphological plasticity and calcium dynamics in dendrites

Area of Science:

• Neuroscience
• Cell Biology
• Microscopy

Background:

• The intricate relationship between nervous system structure and function necessitates simultaneous high-resolution imaging.
• Current methods often lack the spatio-temporal resolution to capture both neuronal morphology and activity concurrently.

Purpose of the Study:

• To develop and present a novel two-step imaging strategy.
• To enable simultaneous visualization of neuronal structure, including dendritic arborization, and calcium dynamics.

Main Methods:

• Utilizing ballistic delivery or single-cell electroporation for fluorescent calcium indicator and red fluorescent dye loading.
• Employing dual-wavelength wide-field or confocal fluorescence microscopy for simultaneous imaging.
• Applying the technique to both developing and mature neurons in live tissue.

Main Results:

• Successfully visualized entire dendritic arborization alongside calcium dynamics.
• Enabled concurrent imaging of structural plasticity (filopodia, spines) and neuronal activity.
• Demonstrated applicability to both developing and mature neurons.

Conclusions:

• The described two-step strategy provides a robust method for high spatio-temporal resolution imaging of neuronal structure and function.
• This technique facilitates the study of the structure-function relationship in the nervous system.
• The protocol requires approximately 2 hours for cell labeling and imaging setup.