Live cell imaging & Tracking
Track and image life cells
Working with living cells is just that bit harder. Huygens offers many solution that make your life easier and improve the quality of your scientific work.
Mitochondrial dysfunction in senescent fibroblasts. 20x air (0.7 NA) widefield image captured on a Leica upright of living human fibroblast cells stained to show the mitochondrial network (green) and it's activity (red) and cell nuclei (blue, using mitotracker green, TMRM and Hoechst respectively). Z stack fluorescent channels were deconvolved in Huygens Essential and rendered using SFP. Individual cells can be seen to have varying levels of energy production (red intensity), as well as intracellular variation between individual mitochondrial networks. Image kindly provided by Dr. Glyn Nelson, University of Newcastle upon Tyne, United Kingdom
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Protocol: less phototoxicity with LIVE deconvolutionLive cell imaging benefits from all the advantages that HUYGENS DECONVOLUTION can offer. One of those advantages is that you need only few photons to reliably restore the underlying object, keeping your cells healthy and allowing you to measure longer or faster. Check out this protocol:
- Load your data. All major microscopy file formats are automatically recognised and read in correctly.
- Stabilize your data - use Huygens Object Stabilizer to correct for any stage drift or cellular movement. In case of a z-stack, stage drift will cause a displacement of the z-planes. Huygens Stabilizer corrects for this as well.
- Deconvolve your data - automatically using Huygens Express or have more control using the Huygens Deconvolution wizard. Works for all major microscopy types.
- Note: Do you have multiple time series? The batch processor automates all these steps for you!
- Visualize your data using Huygens Movie Maker or one of our renderers. Additionally, you can look at your data from any angle and speed-up or slow down your time series.
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. Recorded by Christian Junker M.Sc., Institute of Biophysics, University of Saarland, Germany.
Huygens Object StabilizerHuygens Object Stabilizer can measure and correct for any type of sample movement that is not desired, such as stage drift or cell movement. Huygens Object Stabilizer can stabilize 2D and 3D time series, movement in x, y and z, axial rotation, and also mis-alignment of slices within a 3D stack. Both the measurement and subsequent stabilization are done in 3D and at sub-pixel level.
Time acquisition of this widefield image suffered from a persistent drift and sudden bumb of the stage, preventing proper tracking of the red lysosomes/endosomes. Both stage issues were corrected with Huygens Object Stabilizer. Image shows a mouse embryonic fibroblast (MEF) in which Fascin1 (protein binding to actin, green) and Lamp1 (lysosomal membrane protein 1, red) have been transfected. Courtesy of Prof. Fumio Matsumura, Department of Molecular Biology and Biochemistry, Rutgers University, NJ, USA
Huygens Movie MakerThe Movie Maker is a tool that allows the user to easily create sophisticated animations of multi-channel 3D images using the powerful Huygens visualization tools. Check out our Movie Gallery
Mitochondrial dynamics in human fibroblast. 3D Times series of spinning disk images, deconvolved and rendered in Huygens. Z-stack taken every 20 seconds over half an hour of a live human fibroblast expressing a mitochondrially targeted red fluorescent protein. The image shows part of the mitochondrial network in an old fibroblast's cytoplasm and displays how much movement, fusion and fission happens over time. Courtesy of Dr. Glyn Nelson, University of Newcastle upon Tyne, United Kingdom.
Huygens Object TrackerHuygens Object Tracker tracks the movement of cells and particles in time series images. By manually selecting just a few objects and background regions, the machine learning algorithm in the Object Tracker is sufficiently trained to automatically detect new objects. The included Track Analyzer allows you to filter, edit, and analyze every track. It extracts rotational orientation and movement speed from the tracks and calculates movement speed, diffusion coefficients and Mean-Square-Displacement (MSD). All track data is reported in informative graphs and can be exported to a file.
FUCCI cells tracked for 50 seconds with Huygens Object Tracker. Courtesy of Dr. Richard Wubbolts, Center for Cell Imaging, Faculty of Veterinary Medicine, University of Utrecht, The Netherlands.