Location: Biosciences Room 137

With CLSM, there is often a trade-off between image resolution and speed, spinning disk confocal laser microscopy (SDCLM) overcomes this problem by exploiting the multiplex principle where the sample is illuminated and light detected at multiple points simultaneously. Unlike a conventional laser-scanning confocal microscope with a single pinhole, where a narrow laser beam sequentially scans the sample, in SDCLM an expanded beam illuminates a spinning disk array of microlenses and pinholes which produce multiple focused laser beams which scan across the specimen. Furthermore photomultiplier tubes (PMTs) typically have quantum efficiencies which are rather low, 30-40%, however the SDCLM technique uses a EMCCD camera as a detector that can have a very high QE (Quantum Efficiency) of more than 90%. These differences lead to the systems main advantages: 1) very rapid image acquisition, a 512x512 image every 30ms, so 33 fps, 2) the cooled EMCCD camera allows imaging of very weak signals whilst keeping good signal to noise and 3) The increased sensitivity, which allows the use of lower laser intensities, coupled with the multiple beam scanning method results in a technique which produces very little photobleaching and phototoxicity.

This microscope is good for: High speed, 4D imaging, weak intensity signals, easily bleached fluorophores, FRAP