ENHANCED SCANNING CONTROL IN A CONFOCAL SCANNING LASER MICROSCOPE

A fast and flexible scanning unit, allowing scanning rates of more tha n 1 kHz over regions identified in a specimen, has been developed and evaluated. This scanning unit replaces the original scanning unit in t he Phoibos confocal scanning laser microscope and features full backwa rd compatibility, while at the same time allowing fast and flexible sc anning modes, such as point scanning, line scanning, and scanning alon g user-selected closed curves. The scanning unit uses two galvanometer -mounted mirrors for scanning. A standard procedure for recordings wit h this scanning unit would be to scan an overview image with conventio nal raster scanning to identify a region of interest, mark a point. a Line, or a closed curve over this region, and to start the scanner. An iterating algorithm then calculates the waveforms needed by the scann er to follow the identified curves with pixel precision. With this sca nning unit and its controlling software, experiments demanding lime-re solved recordings within the millisecond range can be performed. Repet ition rates up to > 1 kHz for line scanning and curve scanning, and > 100 kHz for point scanning are obtainable. This allows time-resolved s tudies of fast reactions in living tissue to be performed with the spa tial resolution and signal-to-noise ratio obtainable with a point scan ning confocal microscope.