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Schropp, Martin, Seebacher, Christian and Uhl, Rainer (2017): XL-SIM: Extending Superresolution into Deeper Layers. In: Photonics, Vol. 4, No. 2, 33 [PDF, 6MB]

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Of all 3D-super resolution techniques, structured illumination microscopy (SIM) provides the best compromise with respect to resolution, signal-to-noise ratio (S/N), speed and cell viability. Its ability to achieve double resolution in all three dimensions enables resolving 3D-volumes almost 10x smaller than with a normal light microscope. Its major drawback is noise contained in the out-of-focus-signal, which-unlike the out-of-focus signal itself-cannot be removed mathematically. The resulting "noise-pollution" grows bigger the more light is removed, thus rendering thicker biological samples unsuitable for SIM. By using a slit confocal pattern, we employ optical means to suppress out-of-focus light before its noise can spoil SIM mathematics. This not only increases tissue penetration considerably, but also provides a better S/N performance and an improved confocality. The SIM pattern we employ is no line grid, but a two-dimensional hexagonal structure, which makes pattern rotation between image acquisitions obsolete and thus simplifies image acquisition and yields more robust fit parameters for SIM.

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