Align your multiview Light Sheet images using interactive scenes and real-time visual feedback
Common in Light Sheet Fluorescence Microscopy (LSFM) is the acquisition of multiple (opposing or rotational) views and to fuse these to compensate for light absorption and scattering issues. Typically, interest points (e.g. beads) are used to facilitate this fusion process. Huygens Fuser does not require the use of beads. Instead, interactive scenes allow easy positioning of each sample view, further optimized by advanced correlation algorithms. Using real-time visual feedback of all projected views, you’re in full control over the alignment process!
As the next generation Fusion and Deconvolution Wizard, Huygens versatile Fuser is extremely easy to use and suited for all LSFM data with its additional expert fusion settings, tailored deconvolution for all light sheet types, and CPU + GPU support for efficient computational usage.
Optimal interactive positioning of each view and direct visual feedback makes fusion easy, efficient, and fast.
Specific deconvolution (see picture) and fusion settings for various light sheet setups with extensive help information allow optimal image restoration.
Huygens unique Light Sheet variable PSF modeling ensures high-quality LSFM deconvolution.
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Fusion of various LSFM data
Stripes and shading problems are well-known imaging issues in light sheet microscopy. To minimize these, the sample is typically rotated and imaged from different directions. These different views then need to be fused into one single superior image to have all parts of the specimen imaged optimally. Huygens FUSER is extremely versitale as it helps you with the alignment and fusion of multiview 2D-5D (incl. multi-channel and time) images, which can be aquired with any type of Light Sheet microscope and from any imaging direction.
Two examples of Zeiss Z1 Light Sheet datasets deconvolved and fused with the Huygens FUSER. The first dataset is from a Drosophila brain acquired at 360 degrees rotation (45 degrees steps), and the second set is from a chicken embryo imaged from two opposing sides. Courtesy of Prof. Christophe Marcelle, Mrs. Marie Julie Dejardin (INMG) & Dr. Denis Ressnikoff (CIQLE), Université Lyon 1, France.
Huygens FUSER main window shows on the left previews of each image as maximum intensity projections along the selected axis of rotation. Selected views for fusion are displayed in different customizable colors, and each view can be activated, manually placed, and aligned on top of the others within the center scene by simply using the mouse cursor. After initial setting of the angles and approximate relative shifts, Huygens FUSER fine-tunes the alignment of each view. The FUSER permits full control over the alignment by providing real-time visual feedback during the fusion process. Expert crop, fusion, and sampling settings with detailed explanations are available to cover a broad range of needs, like getting a fused image fast at rough voxel grid size using limited computational resouces, or obtaining at higher computational costs the finest voxel grid size and highest possible resolution.
The Huygens Fuser Window showing multiple views as maximum intensity projections which can be manually aligned before the actual fusion process fine-tunes the alignment. Courtesy of Dr. Denis Ressnikoff from the University Claude Bernard Lyon 1, France.
Light Sheet Deconvolution & Fusion in Huygens
Similar to our previous Huygens Fusion & Deconvolution Wizard, the new Huygens FUSER has the option to deconvolve and fuse Light Sheet images within one single workflow. Several different light sheet setups are supported which include gaussian, high fill factor, scanning and lattice-based systems. By selecting parameter templates customized for your specific LSFM data, both the deconvolution with a light sheet variable PSF and the fusion are easily and reproducibly executed. Huygens unique set of additional restoration options even allow you to correct for additional imaging artefacts like hot&cold pixels, crosstalk (bleedthrough), chromatic aberration, and drift.
Maximum Intensity Projection of a raw (left) and deconvolved (right) 3D image from mouse blastocysts acquired with a Leica Digital Light Sheet microscope. Deconvolution was performed with the CMLE algorithm and the new Huygens module for calculating the theoretical Light Sheet point-spread-function. Courtesy of Dr. Marc Duque Ramirez and Dr. Ritsuya Niwayama (Hiiragi group) and Dr. Stefan Terjung (ALMF) from the EMBL Heidelberg, Germany.
Available for Huygens Essential & ProfessionalThe Huygens Crosstalk Corrector option can be tested with a free test license. The product is commercially available as a licensed option for Huygens Essential, Professional.
Why wait? Try out this option by downloading Huygens and request a test license, or receive pricing information.
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