Parallel confocal detection of single molecules in real time

The confocal detection principle is extended to a highly parallel optical system that continuously analyzes thousands of concurrent sample locations. This is achieved through the use of a holographic laser illumination multiplexer combined with a confocal pinhole array before a prism dispersive element used to provide spectroscopic information from each confocal volume. The system is demonstrated to detect and identify single fluorescent molecules from each of several thousand independent confocal volumes in real time. © 2008 Optical Society of America OCIS codes: 180.1790, 170.2520, 350.4238, 170.6280, 310.6628, 300.2530 Confocal microscopy is a powerful tool for the study of optical phenomena in diffraction-limited observa- We present here a system that provides several thousand continuously monitored confocal observation volumes with single-molecule sensitivity and spectroscopic resolution. In this syste