Light Sheet Fluorescence Microscopy combine fast 3D image acquisition with optical-sectioning by focusing with an excitation objective a thin laser light-sheet into the specimen. This light sheet is perpendicularly positioned with respect to the objective and detector which collect the emitted fluorescent signal as a 2D image. Typically, the specimen is moved along the optical axis to record a three dimensional stack. Multiple stacks can be acquired from different angles ad aligned to account for possible light loss/shading effects.
The Light Sheet technique is very well suited for imaging living specimens since the illumination is restricted to the focal plane which minimizes photodamage and improves contrast. Also no point-scanning is needed shortening the acquisition time significantly.
The Huygens Light Sheet optical option allows deconvolution of large data sets using the robust Huygens deconvolution algorithms. Huygens Light Sheet deconvolution reduces noise and blurring, and takes depth-dependent spherical aberration correction into account. More information on how to deconvolve Light Sheet images in Huygens can be found on Light Sheet deconvolution.
- Improve the quality and resolution of your Light Sheet images using state of the art deconvolution methods.
- PSF adapted to local light sheet thickness.
- Quickly process very large data sets.
- GPU support
- Visualize and quantitatively analyse your data after deconvolution.
Object Stabilizer it is possible to correct for skewing within a Light Sheet image. This is needed in some Light Sheet Fluorescence Microscopy setups, since the excitation and emission axis are oriented at an oblique angle relative to the sample stage. For more information please see the Huygens Object Stabilizer.
Huygens deconvolution of high resolution LIght Sheet data. GFP labeled yolk granules in a C. elegans one-cell stage embryo before (left) and after deconvolution with the CMLE algorithm using a theoretical Light Sheet point-spread-function. SPIM/Light Sheet raw data used by permission from Dr. Uros Krzic, Dr. Lars Hufnagel, and Dr. Yury Belyaev, European Molecular Biology Laboratory, Heidelberg.
Maximum Intensity Projection of a raw (left) and deconvolved (right) 3D image from mouse blastocysts acquired with a 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.