fulltext28 - Multifunctionalized Biocompatible Microspheres...

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Multifunctionalized Biocompatible Microspheres for Sensing R OSARIO M. S´ ANCHEZ -M ART ´ IN ,L OIS A LEXANDER , AND M ARK B RADLEY School of Chemistry, University of Edinburgh, Edinburgh, United Kingdom We present our achievements in the synthesis and application of multifunctionalized, cross-linked, polystyrene microspheres, which remarkably and quite generally are taken up by all cell types studied to date. Importantly, the nature of these synthetic beads allows multistep solid-phase chemistry and the ability to bind essentially any molecule/sensor/nucleic acid to them. These mi- crospheres have a number of advantages over other cellular delivery approaches. First, a diverse range of compounds can be attached to the microspheres, and we have demonstrated that these materials are effectively delivered into cells; second, the cellular cargo can be modulated through modification of bead loading; third, they are large enough to be visualized using standard mi- croscopy techniques; and fourth, their cargos are not diluted within the cell. Populations of cells containing beads can be readily sorted from other cells for subsequent analysis with very high, but controllable, uptake rates, which can be modulated through modulation of the bead size and incubation time. Recent work has focused on the development of these microspheres as sensors for intracellular calcium and pH. Key words: cellular uptake; delivery system; multifunctionalized microspheres; in vitro sensing; fluorescent probes Introduction Biological membranes are natural barriers isolat- ing the content of cells from the extracellular medium and are also responsible for compartmentalization inside the cell. Throughout the membranes various proteins are distributed that are responsible for the controlled efflux/influx of solutes from cells and cel- lular organelles, regulating intracellular concentra- tions of ions and small molecules according to the cells’ needs as well as being otherwise impermeable to most molecules. The introduction of membrane- impermeable molecules into mammalian cells has been an invaluable tool for the study of intracellular processes and has helped hugely in gaining an under- standing of molecular events at the cellular and sub- cellular level. It is of particular interest, with respect to the development of drugs, to understand where their targets are located within host cells. Cellular entry is important for the delivery of potential drugs, small molecule modulators, and sensors. To this end, numer- ous techniques have been developed for the delivery of Address for correspondence: Rosario M. S´anchez-Mart´ ın, School of Chemistry, University of Edinburgh, Joseph Black Building, West Mains Road, Edinburgh EH9 3JJ, UK. rsmartin@staffmail.ed.ac.uk
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fulltext28 - Multifunctionalized Biocompatible Microspheres...

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