Huygens STED Deconvolution Increases Signal-to-Noise and Image Resolution towards 22 nm

November 28, 2013

STED microscopy has proven to be a valuable super-resolution technique, resolving objects that are smaller than the diffraction-limited resolution. Deconvolution of STED images with Huygens pushes the resolution even further. In a recent publication (see link below), we demonstrate that Huygens offers a two-fold improvements of STED images in X, Y, and Z resolution, and increases signal-to-noise ratios eight times. The presented data also shows that a lateral resolution increase from 50 nm towards 22 nm was obtained by applying Huygens deconvolution on (biological) gated-STED images. Furthermore, we describe that stabilization of 3D STED images is essential for optimal deconvolution, as it corrects for lateral drift which would normally distort the structure of the STED PSF.

Overall, the significant SNR increase that can be gained with Huygens deconvolution allow researchers to re-design their (live cell) imaging experiments using shorter exposure times, and minimizing bleaching and photo-toxicity issues.

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Schoonderwoert V, Dijkstra R, Luckinavicius G, Kobler O and van der Voort H:
Huygens STED Deconvolution Increases Signal-to-Noise and Image Resolution towards 22 nm

Microscopy Today 21 (6): 38–44 (2013); doi:10.1017/S1551929513001089

Figure: Deconvolution at work. STED image parameters are automatically read by Huygens and can also be extracted from bead images using the PSF Distiller wizard. The Deconvolution Wizard supports histogram inspection, automatic stabilization, background subtraction, and parameter optimization. Results can be easily compared with the Twin Slicer.

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