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Photonics 2017, 4(2), 33; doi:10.3390/photonics4020033

XL-SIM: Extending Superresolution into Deeper Layers

1
Bio Imaging Zentrum der Ludwig-Maximilians-Universität München, 82152 Martinsried, Germany;
2
TILL I.D. GmbH, 82152 Planegg/Martinsried, Germany
*
Authors to whom correspondence should be addressed.
Received: 14 March 2017 / Revised: 7 April 2017 / Accepted: 13 April 2017 / Published: 20 April 2017
(This article belongs to the Special Issue Superresolution Optical Microscopy)
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Abstract

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 10× 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. View Full-Text
Keywords: structured illumination; confocal microscopy; line-confocal microscopy; 3D-super resolution; hexagonal SIM structured illumination; confocal microscopy; line-confocal microscopy; 3D-super resolution; hexagonal SIM
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This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. (CC BY 4.0).

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Schropp, M.; Seebacher, C.; Uhl, R. XL-SIM: Extending Superresolution into Deeper Layers. Photonics 2017, 4, 33.

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