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Laser-Guided Assembly of Heterotypic Three-Dimensional Living Cell Microarrays

G. M. Akselrod ^*, W. Timp , U. Mirsaidov ^*, Q. Zhao ^*, C. Li , R. Timp ^*, K. Timp ^*, P. Matsudaira  and G. Timp ^*

^* Beckman Institute, University of Illinois at Urbana-Champaign, Urbana, Illinois 61801; and Whitehead Institute, Massachusetts Institute of Technology, Cambridge, Massachusetts

Correspondence: Address reprint requests to Greg L. Timp, University of Illinois at Urbana-Champaign, Dept. of Electrical and Computer Engineering, 3311 Beckman Institute, 405 N. Mathews Ave. MC-251, Urbana, IL 61801. Tel.: 217-244-9629; E-mail: gtimp{at}uiuc.edu.

We have assembled three-dimensional heterotypic networks of living cells in hydrogel without loss of viability using arrays of time-multiplexed, holographic optical traps. The hierarchical control of the cell positions is achieved with, to our knowledge, unprecedented submicron precision, resulting in arrays with an intercell separation <400 nm. In particular, we have assembled networks of Swiss 3T3 fibroblasts surrounded by a ring of bacteria. We have also demonstrated the ability to manipulate hundreds of Pseudomonas aeruginosa simultaneously into two- and three-dimensional arrays with a time-averaged power <2 mW per trap. This is the first time to our knowledge that living cell arrays of such complexity have been synthesized, and it represents a milestone in synthetic biology and tissue engineering.