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Confocal Microscopy Unit

Fluorescence microscopy has become the major tool inmodern biology. Modern microscopy is no longer simply a matter of excellent still images, multiple colours or high optical resolution. Today, scientists want to record brilliant, high-resolution images to illustrate morphological features of fixed or slowly moving live samples, and on the other hand, we demand high-speed dynamic processes occurring in live cells are captured in fast time-course studies, from which significant quantitative data are extracted. These apparently opposing requirements have previously required two classes of systems- those that are optimised for morphological image acquisition, or those designed for high-speed live cell imaging and data generation. The development of new technologies by Leica Microsystems has, for the first time, perfectly merged these two worlds into one.

The new Leica SP5 resonant scanning confocal microscope, located in the Wales Heart Research Institute, combines a true confocal scanner for excellent imaging quality with a resonant scanner technology for high speed imaging (512 x 512 pixels at 25 frames per second, 512 x 32 pixels at 250 frames per second and xt (line) scanning at 16,000 per second).  The growing variety of available fluorochromes featuring different excitation and emission properties has caused a huge demand for new fluorescent filters and dichroics. The revolutionary AOBSä (Acousto Optical Beam Splitter) technology replaces expensive, optically restrictive filter blocks with a switching ‘valve’ for light (actually an acousto-optical crystal, known as a tunable deflection device) and eliminates the chromatic aberration and low transmission inherent in dichroic-based optics. Consequently, AOBS permits a completely tunable spectral band selection for emission, and with near total transmission of emitted photons to ultra-sensitive detectors, it means that images are brighter, crisper and acquired in incredible detail. Furthermore, since the illumination time is so short, there is negligible photodamage to the sample- an important requirement in longer time-lapse experiments. The system accomplishes the most demanding requirements in recent multi-fluorescence research by groundbreaking new technologies to maximise spectral and multi-channel performance.

Our new high-specification system, comprising four lasers (Red-Orange-Yellow-Blue, offering 8 excitation lines), a fully automated state-of-the-art digital microscope, and the capacity to simultaneously capture 4 channels (3 fluorescence channels and 1 dedicated to differential interference contrast (DIC) / brightfield imaging) has been developed in full consideration for future software and hardware upgrades that will be driven by end-user requirements. The incredible imaging power inherent in this new system is tamed by flexible, easy-to-use software (freeware versions of the software can be downloaded onto your individual PCs to enable image analysis wherever, and whenever, you choose) meaning that users can do their own experiments with no prior knowledge of confocal microscopy and following minimal training. Regular in-house training sessions will be available to new and existing users.

Department of Cardiology, Wales Heart Research Institute, Cardiff University School of Medicine, Heath Park, Cardiff, UK CF14 4XN