STED
™Huygens STED Deconvolution Software
Delivers an unprecedented two-fold enhancement in signal and an eight-fold increase in resolution
2D and 3D STED microscopy have proven to be valuable super-resolution techniques for resolving objects smaller than the diffraction-limit, and Huygens' leading STED deconvolution pushes this even further. Huygens can successfully deconvolve all types of STED (STimulated Emission Depletion microscopy) images. Its unsurpassed STED deconvolution provides a two-fold improvement in X, Y, and Z resolution and boosts contrast, with SNR increasing by a factor of eight. A Full Width at Half Maximum (FWHM) resolution of 22nm has been achieved with Huygens STED deconvolution (see our Microscopy Today article). To ensure optimal results, the Huygens STED option includes an integrated STED stabilizer that effortlessly corrects for thermal drift. Note that support for multi-detector STED images requires the Array Detector STED option.
Image description
Primary hippocampal neurons with cytoskeleton proteins labeled (magenta, alpha-Adducin, Abberior STAR 635P and green, ßII spectrin, Alexa 594). Imaged with Abberior Instruments’ STEDYCON, and deconvolved using Huygens.
Reach 22nm Resolution
Improve image resolution, contrast and SNR effectively, while automatically correcting for bleaching, thermal drift and spherical aberration.
Most advanced Algorithms
Use Huygens advanced Maximum Likelihood Estimation algorithms with full GPU acceleration.
All Microscope Brands
All STED systems and their File Formats are supported. Exchange data effortlessly using our links with the Abberior and Leica software.
STED + Huygens Deconvolved
Z-stack MIP projection showing nuclear pore complex protein (red, Abberior STAR RED) and peroxisomes (cyan, Alexa 594) in mammalian cells. Imaged with Abberior Instruments’ STEDYCON and deconvolved with Huygens.
Testimonials
I have been a Huygens user for 8-9 years now and the support provided by you guys are fantastic!
Dr. Priyam Banerjee, image analysis researcher and instructor, MD Anderson Cancer Center, USA.
I've been working with Huygens for couple of years now, and I can say that it's a really powerful tool. SVI have done a great job at developing a software package that is both flexible and user friendly.
Dr. Stefan Stanciu, researcher at the Center for Microscopy-Microanalysis and Information Processing, UPB, Romania.
STED 2D and 3D deconvolution
Huygens STED option deconvolves images acquired using 2D and 3D STED depletion settings. Acquisition with 3D STED depletion is used to enhance the resolution in both the XY and Z direction. To accomplish this the STED beam is split in two, illuminating the sample in both the lateral and axial direction. Huygens STED option includes a specific parameter to address this axial & lateral depletion, which directly influences the aspect ratio of the 3D STED Point Spread Function (PSF) and with that, the extra resolution improvement in X, Y and Z that can be achieved with deconvolution.
Image description
Note the isotropic resolution of the microtubular skeleton of a Marine Dinoflagellate Amphidinium. This 3D STED image (with 50% STED 3X) was acquired with a Leica TCS SP8 STED-3X system, and deconvolved and visualized with the Huygens software. Image provided by Elisa Berdalet, CSIC Institute of Marine Sciences and Timo Zimmermann, Center for Genomic Regulation, Barcelona.
Array Detector STED (Add-on)
The Matrix detector from Abberior uses a grid of 23 detectors to collect more light in confocal images and to distinguish better between background and true signal in STED Images. With the Array Detector STED option as an add-on Huygens has the ability to: read in the 23 detector data perfectly, assess the quality of the alignment in the Array Detector Quality Control tool and optimally deconvolve this multidetector data using a PSF that takes the offsets of the 23 detectors into account. This option contains optimized workflows for both Confocal and STED images.
Image description
MIP projection showing nuclear pore complex protein (Inferno color map, Abberior STAR RED). Imaged with Abberior Instruments’ FACILITY line with a Matrix Detector and 100% 3D STED. Deconvolved with Huygens Array Detector STED option. Raw image provided by Chi Leung So, PTC Internetional Ltd. Hongkong.
Raw Matrix STED
Huygens deconvolution for Abberior and Leica microscopes
All STED microscopy images can be improved with the Huygens software. Huygens also reads the metadata contained within the Leica STED, and the Abberior STEDYCON and Facility line file formats automatically. Upon file reading, a metadata fidelity check is performed to assure optimal deconvolution results. Processing and deconvolution of Abberior Matrix STED requires the Array Detector STED add-on (see above). A special Abberior image feeder for the two Abberior microscope systems enables researchers to transfer images directly from the Abberior's acquisition software to Huygens Professional with a single click. A similar link exists also between the Leica LAS X software and Huygens.Use in research
Anan Chen et al., A coronaviral pore-replicase complex links RNA synthesis and export from double-membrane vesicles. STED images were deconvolved with Huygens. Sci. Adv.10 (2024)
Rodrigues-Oliveira T, Wollweber F et al., Actin cytoskeleton and complex cell architecture in an Asgard archaeon. STED and Airyscan images were deconvolved with Huygens. Nature 613: 332–339 (2023)
Spahn C, Grimm JB, Lavis LD et al., Whole-Cell, 3D, and Multicolor STED Imaging with Exchangeable Fluorophores. STED images were deconvolved with Huygens. Nano Letters 19, 500-505 (2019)
For more, see Scientific PublicationsRelated products
STED 3D stacks often suffer from misalignment of slices due to thermal fluctuations. Therefore, the Object Stabilizer is included in STED deconvolution. The thermal drift correction is also integrated into the specialized STED option for the Huygens PSF Distiller. A distilled PSF from bead images improves the deconvolution results even further and can be used to obtain the STED saturation factor.
Object Stabilizer PSF Distiller
More information
Introduction to deconvolution Huygens Deconvolution software Deconvolution images
Huygens STED Deconvolution Gallery
STED + Huygens Deconvolved
STED140ROR nanorulers (GATTAquant). 70 nm distance between red and white sites. Imaged with Abberior Instruments’ STEDYCON and deconvolved with Huygens Professional.
Gated STED single plane image and confocal data of a centriole stained with primary and secondary antibody conjugated to Chromeo488. Raw and Huygens deconvolved images are shown. The intensity profile was fitted to Lorentz distribution. Image kindly provided by Dr. Grazvydas Lukinavicius, EPFL, Lausanne, Switzerland.
STED + Huygens Deconvolved
Hela cells with actin cytoskeleton labelled (Phalloidin, Abberior STAR Red). Imaged with Abberior Instruments’ STEDYCON and deconvolved with Huygens Professional.
STED + Huygens Deconvolved
Hela cells with actin cytoskeleton labelled (Phalloidin, Abberior STAR Red). Imaged with Abberior Instruments’ STEDYCON and deconvolved with Huygens Professional.
Pre-synaptic marker (Atto647n on 2nd antibody) within the neuromuscular junction of Drosophila melanogaster larvae shows holes in flower structures that were deconvolved with Huygens (picture in the middle), and that can not be identified in the raw Leica STED image (left). Raw pulsed-STED image was first stabilized and then deconvolved with Huygens. Image kindly provided by Oliver Kobler, Ulrich Thomas and Werner Zuschratter, CNI, Leibniz Institute Magdeburg, Germany.
See more: Images in the field of Neurosciences
Confocal and STED image of chromosomes with histon proteins before and after deconvolution. Combined image of chromosomes (confocal in green) and histon protein (STED in red) image before and after deconvolution. More contrast and resolution can be observed after deconvolution of both the Leica confocal and STED channel. The image shows an enlargement of the red channel. Image kindly provided by Dr. Juraj Kabat, Biological Imaging Facility, NIH/NIAID, Bethesda, USA.
See more: Images in the field of Cell Biology
This Huygens deconvolved image shows a IsoSTED image of tubulin labeled with a small molecule probe, and imaged on an Abberior STED expert line microscope. Confocal and 3D STED images were deconvolved using Huygens software. Movie generated using Huygens MIP renderer. Image kindly provided by Dr. Grazvydas Lukinavicius, MPI for Biophysical Chemistry - Department of NanoBiophotonics, Max Planck Institute, Germany.
See more: Images in the field of Cell Biology
Leica pulsed-STED image taken from a neuronal cell-culture model (differentiated SH-SY5Y cells). Cells were fixed and stained with an anti-tubulin primary antibody and anti-rabbit-Atto647N secondary antibody. Inset shows a magnification of one specific area. Image kindly provided by Dr. Heinz-Georg Jahnke/Prof. Dr. Andrea A. Robitzki, Molecular biological-biochemical Processing Technology, Biotechnological-Biomedical Center, Deutscher Platz 5, 04103 Leipzig, Germany. Funded by the Free State of Saxony and the European Union (SMWK/EFRE).
See more: Images in the field of Cell Biology
Both the raw Leica STED image and after subsequent Huygens deconvolution show the two tangled X chromosomes separately at bottom, whereas they can not be separated by regular confocal microscopy. Image kindly provided by Dr. Juraj Kabat, Biological Imaging Facility, NIH/NIAID, Bethesda, USA.
See more: Images in the field of Cell Biology