title: Controlled single-cell deposition and patterning by highly flexible hollow cantilevers.
abstract: Single-cell patterning represents a key approach to decouple and better understand the role and mechanisms of individual cells of a given population. In particular, the bottom-up approach of engineering neuronal circuits with a controlled topology holds immense promises to perceive the relationships between connectivity and function. In order to accommodate these efforts, highly flexible SU-8 cantilevers with integrated microchannels have been fabricated for both additive and subtractive patterning. By directly squeezing out single cells onto adhesive surfaces, controlled deposition with a spatial accuracy of 5 μm could be achieved, while subtractive patterning has been realized by selective removal of targeted single cells. Complex cell patterns were created on substrates pre-patterned with cell-adhesive and repulsive areas, preserving the original pattern geometry for long-term studies. For example, a circular loop with a diameter of 530 μm has been realized using primary hippocampal neurons, which were fully connected to their respective neighbors along the loop. Using the same cantilevers, the versatility of the technique has also been demonstrated via in situ modification of already mature neuronal cultures by both detaching individual cells of the population and adding fresh ones, incorporating them into the culture.
authors.affiliation: Laboratory of Biosensors and Bioelectronics, Institute for Biomedical Engineering, ETH Zurich, CH-8092 Zurich, Switzerland. demko@biomed.ee.ethz.ch.
authors.forename: Vincent
authors.initials: V
authors.lastname: Martinez
authors.affiliation: Laboratory of Biosensors and Bioelectronics, Institute for Biomedical Engineering, ETH Zurich, CH-8092 Zurich, Switzerland. demko@biomed.ee.ethz.ch.
authors.forename: Csaba
authors.initials: C
authors.lastname: Forró
authors.affiliation: Laboratory of Biosensors and Bioelectronics, Institute for Biomedical Engineering, ETH Zurich, CH-8092 Zurich, Switzerland. demko@biomed.ee.ethz.ch.
authors.forename: Serge
authors.initials: S
authors.lastname: Weydert
authors.affiliation: Laboratory of Biosensors and Bioelectronics, Institute for Biomedical Engineering, ETH Zurich, CH-8092 Zurich, Switzerland. demko@biomed.ee.ethz.ch.
authors.forename: Mathias J
authors.initials: MJ
authors.lastname: Aebersold
authors.affiliation: Laboratory of Biosensors and Bioelectronics, Institute for Biomedical Engineering, ETH Zurich, CH-8092 Zurich, Switzerland. demko@biomed.ee.ethz.ch.
authors.forename: Harald
authors.initials: H
authors.lastname: Dermutz
authors.affiliation: Institute of Microbiology, ETH Zurich, CH-8093 Zurich, Switzerland.
authors.forename: Orane
authors.initials: O
authors.lastname: Guillaume-Gentil
authors.affiliation: Laboratory of Biosensors and Bioelectronics, Institute for Biomedical Engineering, ETH Zurich, CH-8092 Zurich, Switzerland. demko@biomed.ee.ethz.ch.
authors.forename: Tomaso
authors.initials: T
authors.lastname: Zambelli
authors.affiliation: Laboratory of Biosensors and Bioelectronics, Institute for Biomedical Engineering, ETH Zurich, CH-8092 Zurich, Switzerland. demko@biomed.ee.ethz.ch.
authors.forename: János
authors.initials: J
authors.lastname: Vörös
authors.affiliation: Laboratory of Biosensors and Bioelectronics, Institute for Biomedical Engineering, ETH Zurich, CH-8092 Zurich, Switzerland. demko@biomed.ee.ethz.ch.
authors.forename: László
authors.initials: L
authors.lastname: Demkó
mesh_terms: D000268:Adhesiveness; D000818:Animals; D018929:Cell Culture Techniques / Q000295:instrumentation*; D002460:Cell Line; D002469:Cell Separation / Q000295:instrumentation*; D002478:Cells, Cultured; D018914:Cells, Immobilized / Q000166:cytology*; D019295:Computational Biology; D003198:Computer Simulation; D004867:Equipment Design; D005103:Expert Systems; D006624:Hippocampus / Q000166:cytology; D056656:Lab-On-A-Chip Devices; D051379:Mice; D008954:Models, Biological; D032446:Myoblasts / Q000166:cytology; D009474:Neurons / Q000166:cytology; D018583:Pliability; D051381:Rats; D017208:Rats, Wistar; D012441:Saccharomyces cerevisiae / Q000166:cytology / Q000254:growth & development / Q000302:isolation & purification; D059010:Single-Cell Analysis / Q000295:instrumentation*; D000076227:Stereolithography; D013499:Surface Properties