The Leica SP5 laser scanning confocal microscope
The Leica SP5 laser scanning confocal system is used for advanced multi-channel fluorescence and live-cell applications. This system has five lasers generating nine lines for fluorescence excitation (405, 458, 476, 488, 496, 514, 543, 594, 633 nm). The system is equipped with an inverted microscope encased with an incubator for live-cell imaging. An acousto-optical beam splitter allows the investigator to rapidly switch collection parameters without the mechanical movement of dichroic mirrors. Emissions are split into a spectrum using a prism and are collected through five separate tunable emission slits/photomultiplier tubes. A parallel transmitted light image can be collected. The system also contains spectral scanning, three-dimensional acquisition/display and time-lapse capabilities. A programmable stage allows automated ‘return-to-site’ and multiple-field stitching. Currently available objectives on the system are: 63x water immersion (Plan-Apo, 1.2 NA), 63x oil immersion (Plan-Apo, 1.4 NA), 40x oil immersion (Plan-Apo, 1.25 NA), 20x multi-immersion (oil, water, glycerin)( Plan-Apo, 0.7 NA) and a 10x air objective (Plan-Fluotar, 0.3 NA). All objectives (except 10x) are capable of DIC imaging. The SP5 microscope is located in HSW-1142.