Huygens Array Detector Optical Option
Supports Zeiss Airyscan®, Fast Airyscan®, nanoSPAD and others!
Huygens latest Array Detector deconvolution option is the first of its kind to be integrated in a powerful GPU accelerated MLE-algorithm, and is now available in Huygens Suite 18.10. The Array Detector optical option is specifically developed to restore images acquired on multi-detector confocal systems, including the well-received and popular Zeiss® Airyscan (1). Thanks to SVI’s newly developed smart multi-detector algorithms, the restoration of raw Array Detector images with Huygens is very efficient, and allows scientists to obtain reliable super-resolution images in which objects at 100nm distance can be distinguished. More importantly, the unique Huygens Array Detector Super-Sampling mode fullfils a wish common to many microscopist as it allows fast image acquisition with less bleaching and phototoxicity, without compromising the improvement in image contrast and resolution.
Reach 100nm resolution
Recent tests with GattaQuant nanorulers (100, 120, 140 nm) confirm 100nm two-point resolution is easily achievable.
Huygens Super-sampling mode permits faster imaging without compromising resolution and contrast.
Multiple detector layouts are supported, including Zeiss (Fast) Airyscan super-resolution, nanoSPAD detectors, and other generic layouts.
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About array-detector confocal systemsWithin array-detector confocal systems, an array of multiple detectors replaces the conventional pinhole and single detector typical for a regular confocal system. Each individual detector acts as a small pinhole, where all but the central detector are slightly displaced with respect to the original pinhole center. The signal from each detector is used to build up an image, and each image is a slightly shifted version of the central detector image. The total (summed) signal collected by all detectors is comparable to that of a single large pinhole in 'classic' confocal. Consequently, an array-detector system has the advantage of combining the benefit of high-signal of a large pinhole confocal system with the high- resolution aspect of a small pinhole confocal system.
The Huygens Array Detector Optical Option allows you to obtain high resolution and contrast from array detector data, such as that acquired on a Zeiss® Airyscan system. The Airyscan microscope can be used in 4 modes, and Huygens now offers high quality deconvolution for each single mode:
- Standard mode: this is the conventional confocal mode of the Airyscan. For Huygens deconvolution you can use the Confocal Optical Option in Huygens setting the pinhole size at the size of the pinhole used during acquisition.
- Virtual pinhole mode: in this mode you can decide for the size of the pinhole in post-processing. For Huygens deconvolution you can use the Confocal Optical Option in Huygens setting the pinhole size at the size of the pinhole decided in post-processing.
- Super Resolution mode: these datasets include all 32-detector images. The Array-detector confocal option in Huygens includes various modes to intelligently combine the information from these 32-detector datasets and deconvolve data acquired in Airyscan super-resolution mode.
- Fast mode: this mode uses elongated laser excitation in combination with 16 detectors (the central 3 'vertical' detector columns). The fast mode allows for the acquisition of 4 lines simultaneously, thereby speeding up the image acquisition with a factor of 4. The Array-detector confocal option in Huygens includes a special Fast Airyscan mode to optimally process the 16-detector datasets from Fast Airyscan.
Next to supporting the various Zeiss Airyscan modes, the Huygens Array Detector Confocal Optical Option also supports other multi-detector layouts. This includes a nanoSPAD25 detector layout, or other generic mulit-detector layouts.
Reach 100nm two-point super resolutionThe combination of Huygens' unique MLE deconvolution and the information available from the array of detectors, now allow researchers to reach 100nm two-point resolution.
By imaging GATTAquant SIM100B nanorulers, which consist of carefully folded DNA origami with fluorophores spaced at a calibrated distance of 100nm, we can confirm the ability of the Huygens Array detector option to reach a two-point resolving capbility of at least 100nm.
Images obtained with Zeiss Airyscan microscope were kindly provided by Dr. Christoffer Lagerholm (Wolfson Imaging Centre, Weatherall Institute of Molecular Medicine, University of Oxford, UK
Huygens Array Detector deconvolution gallery
With 80nm sampling in X and Y, you can resolve 140nm GattaQuant (2) nanorulers with the special superXY mode. Shown are MIPs. The right image is a sum of the unprocessed raw data from all detectors. Image obtained with Zeiss Airyscan microscope kindly provided by Dr. Christoffer Lagerholm (Wolfson Imaging Centre, Weatherall Institute of Molecular Medicine, University of Oxford, UK
With 80nm sampling in X and Y, you can resolve 140nm GattaQuant (2) nanorulers with the special superXY mode. Shown are MIPs. . Image obtained with Zeiss Airyscan microscope kindly provided by Dr. Christoffer Lagerholm (Wolfson Imaging Centre, Weatherall Institute of Molecular Medicine, University of Oxford, UK
Huygens’ unique MLE algorithm - high quality and reliabilityThe Array Detector confocal option in Huygens is able to combine multiple detector images in a classic maximum-likelihood estimation (CMLE) framework to yield a super-resolution image with increased contrast and resolution as compared to images from conventional confocal-based system.
Image formation in fluorescence microscopy is well understood, and can be accurately described with well-known mathematical equations. Image restoration is the inverse problem of trying to solve the equation as best as possible. The maximum-likelihood estimation (MLE) approach, as implemented in the Huygens Software, is known to be best approach to accurately solve the image restoration problem in microscopy. For image restoration to be successful, an MLE approach requires so-called 'a priori' information about the image formation process. Fortunately, we have this information at hand as we have knowledge about the imaging system and the microscopy parameters used. The more 'a priori' knowledge you insert in the equation, the better and more reliable the image restoration result will be.
Fast imaging + Fast high-quality Huygens Super-sampling deconvolutionA particularly useful property of array-detector systems, is that the Nyquist criterion is much less strict as compared to confocal. This allows for imaging at larger pixel sizes and thus faster image acquisition. As a consequence, images will be less affected by photo-bleaching and/or can include a much larger field of view than if acquierd with a confocal setup. Deconvolution of Array detector data can be performed with the specifically designed Huygens Super-sampling mode without any trade-off in SNR and while keeping the super-resolution aspect.
In 2016, Carl Zeiss® introduced the Fast Airyscan mode, with which 4 lines can be scanned simultanously. Huygens also offers deconvolution for these Fast Airyscan images.
In the Airyscan Fast mode, 4 lines can be scanned simultanously by adjusting the excitation profile, and by using 16-detectors in the most central columns in the detector Array. The image acquisition can thus be sped up with an additional factor of 4 when imaging in fast-scan mode. Huygens also offers a super-resolution deconvolution mode for Fast Airyscan images, as it can optimally combine the information from all available detectors, to generate a super-resolution result image of unprecended quality for confocal based systems.
Now Available for Huygens Essential/Professional 18.10The new Huygens option supports various Zeiss Airyscan modes (confocal, virtual pinhole, super-res Airyscan, Fast-mode), nanoSPAD detectors, and also generic Array detector profiles and supports z-stacks, multi-channel, time-series or a combination of these dimensions. The option can be tested with a free test license. The new product is commercially available as a licensed option for Huygens Essential and Professional version 18.10
Why wait? Try out this new option in 18.10 by downloading Huygens and request a test license, or receive pricing information.
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1. Carl Zeiss Microscopy GmbH.
2. GATTAquant GmbH. For more information, see the GATTAquant website.