Separating the One From the Many-Microfabricated Arrays for Cell Separations

Abstract

The separation of single or small groups of cells from within a heterogeneous population is a fundamental need in almost all areas of biomedical research. This effort is required in order to obtain unique cells possessing a desired characteristic for genetic studies, cloning, or other applications. Despite recent technological advances, selection and isolation of individual or small groups of live cells from a population remains a signicant challenge. Most live-cell separation methods require that cells be dispersed into a single-cell suspension, but removal of adherent cells from their growth surface may at times be undesirable. A microfabricated cell array platform composed of releasable elements in combination with either a pulsed laser or needle-based punch system has been developed for analyzing, sorting and collecting viable cells from a mixed population while the cells remain adherent to their growth surface. Both noadherent and adherent cells cultured on the array can be analyzed and selected using standard imaging methods. Target cells can then be collected with high viability and efficiently cloned by releasing the polymeric base or cup in which the cell resides. Benefits of this new approach include high cell viability, small sample size requirements, and broad cell selection criteria. Separation properties include fluorescence signals, morphology, and uniquely, time-dependent variables such as the transmission of properties to daughter cells, growth rate or signaling behavior. Mating the microarrays with image cytometry provides a high-throughput tool for selection and isolation of cells for biomedical and pharmaceutical applications.