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Huygens Fuser

Align multiview Light Sheet images using interactive scenes and automated fine-tuning




Common in Light Sheet Fluorescence Microscopy (LSFM) is to acquire multiple (opposing or rotational) views and 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, each view can be optimally positioned by shifting, rotating and scaling it using the interactive scene; advanced correlation algorithms then fine-tune the alignment. Real-time visual feedback during the alignment process gives you full control over the fusion!

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.

Fuser Ioannis
Multiview 3D light sheet data, fused, deconvolved and visualized with Huygens, shows stained Arabidopsis thaliana mature anther. Scale bar: 30 μm. Courtesy of Ioannis Alexopoulos (GI), Ivo Rieu, Mieke Wolters and Jian Xu, Radboud University, The Netherlands.


Interactive interface

FuserInterface Still
Intuitively shift, scale and rotate each view before correlation algorithms fine-tune the alignment


Expert settings

Interactive02
Specific deconvolution and fusion settings for various light sheet setups allow for optimal image restoration.


Improved by deconvolution

Psf
Huygens' unique Light Sheet Deconvolution is integrated to enhance fusion and form high-quality image result.




Testimonials

I tested the lightsheet fusion with 8-view lightsheet stacks of a drosophila ovary. The fusion worked great! What was especially useful was that it did the registration and fusion without fiducial marker beads in the images.

Dr. Benjamin Lopez, facility director NRI-MCDB Microscopy, University of California Santa Barbara, USA.
Deconvolution of lightsheet data is not a trivial task, and also fusion of multiviews is in my experience with other software packages a trial and error process. I was really amazed by the Fusion Wizard in Huygens that actually accomplishes both tasks fast and efficiently

Dr. Marco Marcello, imaging manager Centre for Cell Imaging, University of Liverpool, UK.



Screenshot From 2019 10 03 16 46 10

Interactive interface with automated fine-tuning


The fusion process consists of two main parts: manual approximation followed by automatic optimization. In the interactive Fuser window, the user can approximately align the different views in an intuitive manner by zooming, shifting and rotating them on top of each other. Optional settings, including pixel weighting and cropping and sampling settings, allow for further customization of the fusion.
After the manual approximation, the Fuser uses advanced correlation algorithms to fine-tune the alignment. Huygens' unique Light Sheet Deconvolution is integrated in the fusion pipeline, allowing for fusion based on fine image details and ensuring a high-quality final image.

Image description
The Huygens Fuser window shows maximum intensity projections of the multiple views, which can be manually aligned before the correlation algorithm fine-tunes the alignment. Courtesy of Dr. Denis Ressnikoff, University Claude Bernard Lyon 1, France.



All dimensions, all LSFM types


Huygens Fuser can be used to fuse images acquired with any type of Light Sheet microscope and from any imaging direction. In addition, the Fuser can process data from 2D to 5D (multi-channel 3D time series).

Image description
Two Light Sheet datasets deconvolved and fused with the Huygens Fuser. The top dataset is from a Drosophila brain acquired at 360 degrees rotation (45 degrees steps). The bottom set is from a chicken embryo imaged from two opposing sides. Both sets courtesy of Prof. Christophe Marcelle, Mrs. Marie Julie Dejardin (INMG) and Dr. Denis Ressnikoff (CIQLE), Université Lyon 1, France.
Screenshot From 2019 10 10 17 04 08





Light Sheet Deconvolution & Fusion in Huygens


The Huygens Fuser has the option to deconvolve and fuse Light Sheet images within one single workflow. Several different light sheet setups are supported, including but not limited to gaussian, high fill factor, scanning and lattice-based systems. By selecting parameter templates customized for your specific LSFM system, both the deconvolution and the fusion are easily and reproducibly executed. Huygens unique set of additional restoration options allow you to correct for additional imaging artefacts like hot&cold pixels, crosstalk (bleedthrough), chromatic aberration, and drift.

Video description
Maximum Intensity Projection of a raw (left) and deconvolved (right) 3D light sheet image of a mouse blastocysts. Huygens’ unique theoretical Light Sheet PSF was used for deconvolution. Courtesy of Dr. Marc Duque Ramirez, Dr. Ritsuya Niwayama and Dr. Stefan Terjung, EMBL Heidelberg, Germany.



Use in research

Hötte K, Koch M, Hof L et al., Ultra-thin fluorocarbon foils optimise multiscale imaging of three-dimensional native and optically cleared specimens.
Huygens was used for Light Sheet fusion and deconvolution.
Sci Rep 9, 17292 (2019).

For more, see Scientific Publications

In some Light Sheet setups, the sample is moved horizontally across an obliquely oriented excitation beam. The resulting images are then skewed and should be stabilized before further processing. Light Sheet images can be deconvolved reliably with Huygens' specialized Light Sheet Deconvolution.

Object Stabilizer Light Sheet Deconvolution

More information

Support page
Light sheet deconvolution
Fuser webinar