Controlled single-cell deposition and patterning by highly flexible hollow cantilevers

Area of Science:

• Biotechnology
• Neuroscience
• Materials Science

Background:

• Single-cell patterning is crucial for understanding cellular roles and mechanisms.
• Engineering neuronal circuits with controlled topology offers insights into connectivity and function.

Purpose of the Study:

• To develop a flexible microfabrication technique for precise single-cell patterning.
• To enable both additive and subtractive patterning of neuronal cells for circuit engineering.

Main Methods:

• Fabrication of flexible SU-8 cantilevers with integrated microchannels.
• Controlled deposition of single cells via squeezing onto adhesive surfaces (5 μm accuracy).
• Subtractive patterning through selective removal of single cells.

Main Results:

• Achieved precise single-cell deposition and removal using SU-8 cantilevers.
• Created complex neuronal patterns, including a connected circular loop of hippocampal neurons.
• Demonstrated in situ modification of mature neuronal cultures by adding and removing cells.

Conclusions:

• The developed SU-8 cantilever technique offers high flexibility and spatial accuracy for single-cell patterning.
• This method facilitates the engineering of neuronal circuits with controlled topology.
• The technique supports long-term studies and in situ modifications of neuronal cultures.