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Spinning Disc Confocal Microscopy

Our spinning disc microscope was actually purchased commercially from Andor and is based on an inverted nikon TE2000 microscope. We can excite at 488nm (coherent), 561nm (cobolt) and 633nm (coherent) to obtain fast images of an object. The spinning disc has its name from two fast rotating discs with numerous either holes or microlenses in a spiral arrangement. Each hole-microlens combination gives rise to a tiny confocal spot on the sample which are rapidely moving across the whole sample. With the integration speed of the field detector, usually an EMCCD, it gives an almost confocal image with a fine z resolution at widefield image acquisition speed. The yokogawa CDU10 spinning disc unit together with a z-piezo gives us then a whole volume of the sample at high acquistion rates. In order to minimize the spectral crosstalk, an optosplit unit is attached before the signal is recorded with an andor IXon EMCCD. The system is controlled by the revolution system of the same company. Our experiments on this setup deal mainly with the dynamics inside living cells: mRNA transport, chromosome separation and particle uptake.


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