Encoding microcarriers by spatial selective photobleaching

Bead-based assays on very large numbers of molecules in gene expression studies, drug screening and clinical diagnostics 1 , require the encoding of each of the microspheres according to the particular ligand bound to its surface 2 , 3 . This allows mixing the uniquely encoded microspheres and subjecting them to an assay simultaneously. When a particular microsphere gives a positive reaction, the substance on its surface can be identified by reading the code. Previously reported techniques for colour encoding polymer microspheres 4 , 5 , 6 , 7 , 8 , 9 , 10 only allow for a limited number of unique codes. Graphical encoding methods use metallic particles 11 , 12 , which are rather uncommon in screening applications. Here, we demonstrate a new approach to encode polymer microspheres that are commonly used in screening applications, such as polystyrene microspheres, with a method that provides a virtually unlimited number of unique codes. Patterns can be written in fluorescently dyed microspheres by 'spatial selective photobleaching' and can be identified by confocal microscopy. Such encoded microparticles can find broad application in the collection and analysis of genetic information, high-throughput screening, medical diagnostics and combinatorial chemistry, and can also be used for labelling of consumer goods or as security labels to prevent counterfeiting.