INTRODUCTION TO LASER SCANNING MICROSCOPYLSM 510 Performance Features of the LSM 510B 40-051 e 07/98 3-73.4 Performance Features of the LSM 5103.4.1 Optical and mechanical aspectsThe highly integrated system design makes for the shortest possible optical paths, top-grade opticalprecision and high stability. The compact scanning module can be fitted to an inverted (Axiovert 100 MBP or SP) or upright (Axioplan 2 MOT ) microscope in less than three minutes. On the Axiovert, thescanning module may be mounted either to the base port directly below the microscope or to the sideport.The spectral range available extends from the UV to the IR region.For the VIS (visible-light) Laser Module, the user can select from up to five lasers with wavelengths of633, 568, 543, 514, 488 and 458 nm. The UV Laser Module provides wavelengths of 351 and 364 nm.Coupling of the laser light is through polarization-preserving single-mode optical fibers. One variablebeam collimator each for the UV and visible ranges provides optimum adaptation of the respective laserwavelength to the objective used and, thus, optimum correction for Z aberrations.Acousto-optical tunable filters (AOTF) adjust the necessary brightness for all desired laser lines withinmicroseconds.A monitor diode permanently registers the laser output; it can be used for the on-line checking of theintensity of the exciting light. This check is also possible selectively for the different wavelengths if a lineselection filter is inserted.The four simultaneous image acquisition channels, usable for reflection or fluorescence, and anadditional transmitted-light channel are ideal for the investigation of multiple fluorescence specimens.Separately in each of the four channels, the diameters of the pinholes and their XY positions can beoptimized, and the desired emission filter placed into the beam path, by servo-motor control. In the caseof pinhole VP1, this adjustment also includes positioning along Z. In the simultaneous registration ofmultiple fluorescences, identical optical sections can be obtained in each confocal channel. This is ofimportance, e.g., with the FISH method (fluorescence in-situ hybridization) used for genome analysis incytogenetic studies.The microscope's transmitted-light channel is equipped with a photomultiplier, too. It is thereforepossible to superimpose a multiple fluorescence image on a brightfield, differential interference or phaseimage.A fiber-optic cable connection to external special detectors, such as cooled PMTs or spectrometers, isunder development.In addition to the emission filters for all standard and special applications, available in motor-controlledfilter wheels, the user can easily install his own emission filters in two of the channels.The high-NA C-APOCHROMAT objectives specially developed for the LSM technique reach the physicallimit in resolving power, and can be used throughout the 350...700 nm spectral range with the samehigh quality, producing brilliant images.A two-mirror scanner system, controlled by a digital signal processor (DSP), offers several advantages.The large deflection angle of the scanning mirrors allows a wide area to be scanned. With a 1.25xobjective, the object area scanned is 10 x 10 mm².