Initiating Rapid Neurite Extension and Formation Functional Neuronal Connections

Abstract

Source: M. H. Magdesian et al., Rewiring Neuronal Circuits: A New Method for Fast Neurite Extension and Functional Neuronal Connection, J. Vis. Exp. (2017)

This video demonstrates a technique for the rapid initiation and extension of a neuron and creation of new connections with a target neuron using a microfluidic device and micromanipulation. This could be used for therapies that aim to reconnect neuronal circuits after trauma or in neurodegenerative diseases.

Protocol

1. Standardization of Neuronal Cultures Using Microfluidic Devices: Device Assembly

1. Select a suitable microfluidic device for the desired experiment. To connect neurons within the same population, use the Neuro Devices (Figure 1), and to connect neurons in different populations, use the Co-Culture Devices (Figure 2).
2. Clean and prepare the desired number of sterile coverslips or glass bottom dishes. For best results on plastic surfaces, use 35 mm dishes, on glass, use 25 mm coverslips or 35 mm glass bottom dishes. Select glass thickness based on the imaging system, for instance, 0....

Results

Figure 1: Standardization of Neuronal Cultures using Microfluidic Devices7. (a) Device assembly: when the microfluidic devices are properly assembled on a dry surface all chambers are visible. (b) Cell plating: plate the .......

Materials

List of materials used in this article

Name	Company	Catalog Number	Comments
Neuro devices	Ananda Devices		Commercially available at http:// www.anandadevices.com
No. 1 Glass Coverslip 25 mm Round	Warner Instruments	64-0705
35 mm cell culture dish, NonPyrogenic, Sterile	Corning Inc	430165
95 mm x 15 mm Petri Dish, Slippable Lid, Sterile Polystyrene	Fisherbrand	FB0875714G
Neurobasal Medium	Life Technologies	21103-049	Extracellular solution
B-27 Supplement (50X), serum free	Thermo Fisher Scientific	17504044	Extracellular solution
Pennicilin, Streptomyocin, Glutamine	Thermo Fisher Scientific	11995-065	Extracellular solution
2 - 20 μL Pipettors Aerosol Resistant Tips	Thermo Fisher Scientific	2149P
Dumont Dumostar Tweezers 11 cm	World Precision Instruments	500233
Dissection tools	Braun, Aesculap
Poly-D-lysine Hydrobromide	Sigma-Aldrich	P6407
Micro particles based on polystyrene, 10 μm	Sigma-Aldrich	72986
Horizontal Pipette Puller	Sutter Instruments	Brown-Flaming P-97
Micromanipulators, PCS-5000 Series MC7600R	SD Instruments	MC7600R
1 mL Syringe	BD Luer-Lok	309628
Inverted Microscope	Olympus	IX71
Objective UIS2	Olympus	LUCPLFLN 40X
CCD Camera	Photometrics	Cascade II: 512