The development of microfabrication and microfluidic technologies for biological studies has enabled cell and microorganismal behavior to be examined with increasingly high precision. For example, the use of microfabricated landscapes, microfluidic circuits, and droplet generators have been used to study a diverse range of cell behaviors including microbial evolution, biofilms, chemotaxis, and cellular communication. In general, these techniques rely on essentially stochastic interactions to confine cells in low dimensional volumes. This severely limits abilities to isolate a single targeted cell of choice, which would prove useful for studying particularly challenging problems, for instance, phenotypic diversity of clonal populations or analysis of rare/unculturable microorganisms.We have developed materials and synthetic procedures that begin to address these criteria and demonstrate�under conditions that can maintain cell viability�precise physical confinement of target cells and their progeny within 3D micro-chambers built from protein hydrogels. Isolation of targeted cells is achieved via rapid solidification of protein-based matrices from a biocompatible precursor solutions using multiphoton lithography, an intrinsically 3D laser direct write micro- nanofabrication technique. Cells remain chemically accessible to environmental cues enabling their propagation into well defined, high density populations. We demonstrate this methodology on gram negative (E. coli), gram positive (S. aureus), and eukaryotic (S. cerevisiae) cells. The opportunities to confine viable, single/individual-cells and small populations within user-defined microenvironments afforded by this approach should facilitate the study of cell behaviors across multiple generations (see Anal Chem 2012 84  21:8985-9).

Biocompatible microfabrication of 3D isolation chambers for targeted confinement of individual cells and their progeny

246th National Meeting (2013)

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