What is it about?
We present here a visual protocole of a method based on the combination of fluorescence exclusion, low-roughness multi-compartments microfluidic devices, and finally micropatterning to achieve in vitro measurements of local neuronal volume. The high resolution provided by the device allowed us to measure the local volume of neuronal processes (neurites) and the volume of some specific structures involved in neuronal growth, such as growth cones (GCs).
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Why is it important?
Axonal length and diameter are tightly regulated in vivo, with values specific to each neuronal type. Volume is indeed an important signature of physiological and pathological events in neuronal life, from transient axonal deformation associated to electrical activity to the irreversible neuronal swelling occurring during the asymptomatic phase of neurodegenerative diseases. Here we provide a method to access with high precision the volume and height of neuronal branches and dynamic structures involved in neuronal growth like growth cones.
Perspectives
A possible evolution of this method would be to get rid of this physical confinement, to be replaced by an optical one. The new development of light sheet microscopy might be combined advantageously with FXm in the future.
Dr Sylvain Monnier
Université Claude Bernard Lyon
Read the Original
This page is a summary of: High-resolution Volume Imaging of Neurons by the Use of Fluorescence eXclusion Method and Dedicated Microfluidic Devices, Journal of Visualized Experiments, March 2018, MyJove Corporation,
DOI: 10.3791/56923.
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