Huygens STED deconvolution
STED (Stimulated Emission Depletion) microscopy allows super-resolution imaging in the 50nm range. Unfortunately, this increased optical resolution also leads to a drawback: because many fluorophores are depleted by the depletion laser, this also results in a lower signal (fewer photons) being captured by the detector. Because of the Poisson nature of photon statistics, the signal-to-noise ratio of the resulting image will decrease compared to normal confocal imaging. Therefore the SNR value in STED images will be much lower compared to their confocal counterpart.
Luckily, Huygens deconvolution of STED images offers a great solution. Since 2012, the Huygens software is able to generate a theoretical PSF based on STED microscope parameters. This theoretical PSF can be used in the deconvolution process, which will significantly reduce the noise in the image, and will increase the contrast of up to 10 times. Additionally, the Huygens deconvolution algorithms are able to increase the resolution in the image with a factor of 2 in both lateral and axial direction. See also our Microscopy Today article on Huygens STED deconvolution.
On this page, you find all practical information needed to optimally deconvolve your STED images. A theoretical background of STED can be found on the STED Microscopy page.
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