Imaging Live Samples
Life Happens in 3D
Light-sheet microscopes are powerful tools for live cell imaging that provide unique access to the complex structure and behavior of whole embryos, cell cultures, and organoids. These instruments capture high-resolution images significantly faster, over longer acquisition times, with far less photodamage than other microscopes used in live cell studies. As a result, light-sheet microscopes uniquely enable observation and precise tracking of highly complex and dynamic molecular and cellular events in an almost unlimited range of organisms, particularly during embryonic development and in 3D cell culture applications. These in vivo studies are integral to leading-edge developmental, cell biology, and organoid research.
Embryogenesis and Developmental Biology
For more information about using light-sheet microscopes for developmental biology research, click here:
Drosophila Embryo Development
Transgenic line expressing His2Av-mCherry as fluorescent nuclear reporter. The fruit fly embryo was imaged for almost one complete day (4 × 200 slices every 30 seconds). Imaged on the MuVi SPIM.
Courtesy of:
Lars Hufnagel
European Molecular Biology Laboratory (EMBL)
Heidelberg, Germany
Cell Tracking in Drosophila Embryo Development
Cell tracking created with arivis Vision4D 3D visualization and analysis software. Imaged on the MuVi SPIM.
Courtesy of:
Celia Smits and Stanislav Y. Shvartsman
Department of Molecular Biology
Princeton University, NJ
USA
Zebrafish Development
Zebrafish imaged on the MuVi SPIM. Stitched from 5 stacks in Imaris, each 340 slices, 16 hour/10min. Fish growth can be observed.
Courtesy of:
Prof. Jingxia Liu
Huazhong Agricultural University
China
Zebrafish Embryonic Development
Zebrafish embryo expressing Histone H2A-GFP imaged every 6 min from late gastrula to 15–17 somite stage. Imaged on the MuVi SPIM.
Courtesy of:
Andres Collazo
Caltech, Pasadena, CA, USA
as well as: Course faculty and participants of the 2017 Zebrafish Course
Marine Biological Laboratory (MBL)
Woods Hole, MA, USA
Vascular Development in Zebrafish
From left to right: the video shows a beating Zebrafish heart imaged at 50 frames/sec, followed by Zebrafish blood vessels (magenta) and red blood cells (yellow) and Zebrafish blood flow imaged at 50 frames/sec. Imaged on the MuVi SPIM.
Courtesy of:
Nadia Mercader & Inés Marques
University of Bern
Bern, Switzerland
Drosophila Egg Chambers
Drosophila ovariole stained with phalloidin to label actin (red) found along membranes and the germline ring canals, DAPI (blue) to show the nuclei, and a somatic ring canal marker (green) to label the ring canals in the epithelium.
Courtesy of:
Jasmin Imran Alsous
Schvartsman Lab
Princeton University, Princeton, NJ, USA
3D Imaging of Organoids
Pancreatic Sphere
hESC-derived pancreatic spheres. Imaging on the TruLive3D Imager enables collecting information of several samples in one experiment. Visualization: Imaris (Bitplane)
Courtesy of:
Yung Hae Kim
Graphin-Botton Group, MPI-CBG
Dresden, Germany
Tumorigenesis in Mammary Organoids
Characterization and imaging of stochastic tumorigenesis in mammary organoids. Imaged on the InVi SPIM.
A. Alladin, L. Chaible, L. Garcia del Valle, S. Reither Sabine, M. Loeschinger, M. Wachsmuth, J.K. Hériché, C. Tischer, M, Jechlinger. Tracking cells in epithelial acini by light-sheet microscopy reveals proximity effects in breast cancer initiation. eLife 2020;9:e54066 doi: 10.7554/eLife.54066
3D Cell Culture
3D culture system of human primary cells imaged on the QuVi SPIM.
Courtesy of:
Yassen Abbas
Turco Lab, University of Cambridge
Cambridge, UK
Fixed Astrocyte Spheroid
Spheroid stained with anti-GFAP (Alexa 488) to label astrocytes and anti-Neurofilament200 (Alexa555) to label neurons. Imaged on the InVi SPIM.
Courtesy of:
Markus Bruell
AG Leist, University of Konstanz
Konstanz, Germany
3D Cell Culture
3D culture system of human primary cells imaged on the QuVi SPIM (330µm x 220 µm x 1200µm). Imaged on the QuVi SPIM.
Courtesy of:
Yassen Abbas
Turco Lab, University of Cambridge
Cambridge, UK
3D Imaging of a Spheroid
Spheroid labeled with EGFP and mRFP imaged on the InVi SPIM Lattice Pro. Three illumination patterns were tested for each label: Gaussian beams, Bessel beams, and optical lattices. The optical lattices gave the best results for the EGFP labeling, while the Gaussian beam was optimal for the mRFP labeling.
Courtesy of:
Martin Stöckl
University of Konstanz
Germany
Colonies of Mouse Embryonic Stem Cells
Colonies of mouse embryonic stem cells stably expressing H2B-mCherry and IRFP670 with a membrane-targeting signal. Imaged on the InVi SPIM.
Courtesy of:
Pierre Neveu
European Molecular Biology Laboratory (EMBL)
Heidelberg, Germany
Cell Culture Imaging
Mitosis in HeLa Cells
Mitosis in HeLa cells stained for histone 2B-mCherry (magenta), GFP-tubulin (green) and GFP-tubulin (white, deconvolved).
Imaged on the InVi SPIM Lattice Pro.
Visualization: Imaris (Bitplane).
Courtesy of:
Sabine Reither
European Molecualr Biology Laboratory (EMBL)
Heidelberg, Germany
Sample: HeLa cells (Neumann et al., Nature. 2010 Apr 1;464(7289):721-7)
Mouse Pre-implantation Development
Left: Mouse preimplantation embryos expressing H2B-mCherry. Nuclei tracking from one-cell stage to blastocyst.
Right: Mouse oocytes expressing CENPC-EGFP and H2B-mCherry for kinetochore tracking.
Imaged on the InVi SPIM.
Petr Strnad, et al. (2016). Inverted light-sheet microscope for imaging mouse pre-implantation development. Nature Methods 13, 139-145
Epithelial Cells
Epithelial cell line (BS-C-1) stained for f-actin and chromatin. Image taken with InVi SPIM Lattice Pro.
Courtesy of:
Ulrike Engel
Nikon Imaging Center
University of Heidelberg
Germany
HeLa Cell Culture
HeLa cells expressing GFP and mCherry. Imaged on the InVi SPIM.
Courtesy of:
Tobias A. Knoch
Erasmus MC
Rotterdam, The Netherlands
Neurobiology and Neurodevelopment
Microglia Response to Axonal Damage
The axon of a neuron in the zebrafish brain was selectively dissected by means of IR laser ablation (MuVi SPIM). Thirty minutes after ablation, four microglia reached the damaged axon.
Image taken from:
de Medeiros, G., Kromm, D., Balazs, B. et al. Cell and tissue manipulation with ultrashort infrared laser pulses in light-sheet microscopy. Sci Rep 10, 1942(2020). https://doi.org/10.1038/s41598-019-54349-x
C. elegans Lineage Tracing
The histone marker (H2A::mCherry, purple) and the membrane marker (cnd-1::GFP, green) were used for lineage tracing in C. elegans. Imaged on the QuVi SPIM.
Courtesy of:
Zhirong Bao
The Zhirong Bao Lab
Memorial Sloan Kettering Cancer Center (MSKCC)
Ney York, USA
Microglia Movement
Microglia movement in zebrafish. The vascular system is labeled with a cyan marker and microglia with a yellow one. Imaged on the QuVi SPIM at 2 FPS for 20 min. Two orthogonal views fused and max. project.
Courtesy of:
N. Norlin, F. peri
European Molecular Biology Laboratory (EMBL)
Heidelberg, Germany
Zebrafish Eye
Zebrafish eye imaged on the MuVi SPIM.
Courtesy of:
Anja Machate and Michael Brand
Center for Regenerative Therapies Dresden (CRTD), TU Dresden
Dresden, Germany
Newborn Mouse Cochlea
Hair cells stained for GFP in a newborn mouse cochlea. Imaged at a magnification of 62.5x. Imaged on the InVi SPIM.
Courtesy of:
Raphael Etournay
Genetics and Physiology of Hearing, Institut Pasteur
Paris, France
Live Imaging of Plants
Arabidopsis Root Growth
Transgenic Arabidopsis root expressing nuclear envelope marker imaged with the MuVi SPIM. The comparison of the videos shows the effect of gravity on root growth.
Courtesy of:
Shanjin Huang
Tsinghua University
Beijing, China
Arabidopsis Root
Transgenic Arabidopsis root expressing a membrane marker. Imaged with the InVi SPIM.
Courtesy of:
Alexis Maizel
COS, University of Heidelberg
Heidelberg, Germany
Microalgae
Autofluorescence in microalgae. Imaged on the InVi SPIM.
Other Applications
Mammalian Tumors
Mammalian tumors (red) growing in zebrafish (green). Imaged on the MuVi SPIM
Courtesy of:
Dr. Xi Yao and Dr. Zhangzhao Junjie
Model Animal Research Center of Nanjing University
China
Zebrafish Heart Beating
Zebrafish heart beating imaged on the MuVi SPIM.
Courtesy of:
Prof. Jingxia Liu
Huazhong Agricultural University
China
C. elegans Skeleton Protein Dynamics
Protein dynamics in C. elegans skeleton. Imaged on the InVi SPIM.
Courtesy of:
Dr. Chai Yongping and Dr. Ou Guangshuo
Tsinghua University
China
Daphnia Magna
Auto-fluorescence in Daphnia imaged on the MuVi SPIM.
Courtesy of:
Ellen Decaestecker and Luc De Meester
KU Leuven
Kortrijk, Belgium
