Pipette micromanipulation guides the extension (> 250 µm) of the new filament from population one over the gap and the formation of a connection with the neuronal population two.

 

Connection of two isolated neuronal populations using the co-culture device.

(a) Co-culture device (blue) assembled on a glass coverslip to grow four isolated neuronal populations on one coverslip.

(b) After 14 days in culture, the Co-culture device was removed, exposing the 200 μm gap between neuronal populations.

(c) Schematic representation of the experimental setup shows a zoom in the gap between two isolated neuronal populations (population one above and population two below) as well as the position of the two pipette tips.

(d) By applying negative pressure to a pipette, a PDL-bead adhered to neuronal population one is pulled with the pipette tip, thereby initiating a new filament. By maintaining the negative pressure in the pipette, the PDL-bead-new filament complex (green) can be pulled enabling filament elongation.

(e) Pipette micromanipulation guides the extension (> 250 µm) of the new filament over the gap and the formation of a connection with the neuronal population two. To ensure adhesion of the new filament to neuronal population two, a PDL-bead (red) is positioned (with a second pipette) on top of the extended filament and neuronal population two. Image shows two newly induced filaments guided with micromanipulation to connect two previously isolated neuronal populations in less than 2 h.

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