Huygens Professional Package and 3D rendered with the Huygens Surface Renderer. The movie was created with the Huygens Movie Maker.
The novel Leica TCS SP8 STED 3X system now also allows for super-resolution in the axial (z) direction, while also keeping the conventional super-resolution in lateral (x,y) directions. The special Huygens STED 3D deconvolution module provides significant noise reduction, contrast increase and even more resolution increase for these powerful STED systems.
Specifically notice that in confocal mode (red) the structure is completely blurred out in axial direction due to the confocal diffraction limit, while the STED 3X deconvolved image demonstrates that the fine microtubular structure can be easily resolved thanks to the additional super-resolution in axial direction.
Original dataset, both confocal and STED 3X mode are imaged with a Leica TCS SP8 STED 3X system.
Microtubular staining after methanol fixation using a primary antibody against a Tubulin and an Oregon Green 488 conjugated secondary antibody.
Acknowledgement: Elisa Berdalet, CSIC Institute of Marine Sciences/Timo Zimmermann, Center for Genomic Regulation, Barcelona
Jurkat T-cell making contact with a Raji B-cell, stained for Raji cytosol (FURA-2, blue), T-cell receptor (anti-CD3 Alexa647, green), and Actin (Lifeact-mRFPruby, red).Animated SFP rendering of a deconvolved time series. Original data was imaged using a Zeiss Cell Observer HS system with a 40x Fluor 1.3NA oil lens and deconvolved using Huygens Essential. The original time interval is 1 minute. "We look at the molecular events and redistribution of certain markers for T-cell activation like the TCR (green) and Actin (red). In the time series a ring-like pattern of Actin and a bulls-eye pattern of CD3 can be seen, accumulating at the contact-site (the immunolgical synapse. Both structures gave us a hint that the T-cell was activated properly." Recorded by Christian Junker M.Sc., Institute of Biophysics, University of Saarland, Germany.
SFP Rendering of a cell cluster.Imaged using an inverted Leica TCS SP2 AOBS confocal microscope and deconvolved using Huygens Professional. The original data was recorded by Dr. Nicolas Fete, Laboratory of Stem Cells Dynamics, Swiss Federal Institute of Technology of Lausanne, Switzerland.
Multiple renderer animation made with the new Huygens Movie Maker.The data shows an isolated rat hepatocyte couplet. Recorded by Dr. Permsin Marbet in the lab of Prof. Lukas Landmann, Department of Anatomy, University of Basel, Switzerland.
Huygens Professional. The movie was created with the Surface Renderer in Huygens. It can be observed that in a metaphase cell kinetochores (red) attach to microtubules. Courtesy of Dr. Claudia Florindo, Microscopy Unit, Department of Biomedical and Medical Sciences, University of Algarve, Portugal
SFP renderer and saved as a movie file with Huygens. Courtesy of Barna Dudok, Molecular Neurobiology, Institute of Experimental Medicine, Hungarian Academy of Sciences, Budapest, Hungary.
HeLa cells stably expressing GFP-a-tubulin and mCherry-H2B have been transiently transfected with SNAP-tagged Actin. Cells were stained with appropriate substrate and imaged with a Leica SP5 in resonance scanned mode. Acquired z-stack were deconvolved using Huygens Essential software and represented as a movie generated by the SFP Renderer. Courtesy of Dr. Grazvydas Lukinavicius, Institute of Chemical Sciences and Engineering, École Polytechnique Fédérale de Lausanne (EPFL), Lausanne, Switzerland
chondrocyte within human cartilage was stained with an antibody specific for TSG-6 (Tumour Necrosis Stimulated Gene-6; secondary antibody is conjugated to Alexafluor 555) and probed with biotinylated Hyaluronan-Binding protein (b-HABP) to identify Hyaluronan (HA). 3D time images were deconvolved and subjected to the Surface Renderer in Huygens. Courtesy of Dr. Sheona Drummond, Wellcome Trust Centre for Cell Matrix Research, Faculty of Life Sciences, University of Manchester, United Kingdom.
Recruitment of DsRed2-PKD1 from the cytosol to the immune synapse occurs after the Jurkat cell recognizes and binds to the Raji B lymphocyte. This recruitment is much more obvious after deconvolution. Courtesy of Dr. Manuel Izquierdo, CSIC-Universidad Autónoma de Madrid, Spain.
Tetrahymena was stained for centrin (red) and Mob1-GFP (green). Stacks were acquired with a Delta Vision microscope and images were subsequently deconvolved with Huygens. The movie was created with the Huygens Surface Renderer and Movie Maker. Centrin staining is present in the cilia of Tetrahymena, and Mob1 staining is found in the apical region of this organism. Courtesy of Dr. Claudia Florindo, Microscopy Unit, Department of Biomedical and Medical Sciences, University of Algarve, Portugal