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Light-sheet fluorescence microscopy has become the method of choice for non-invasive imaging of a variety of live and cleared biological samples ranging from subcellular structures to cells, tissues and whole embryos, e.g. Drosophila and Zebrafish.

Among other features, the significantly reduced photodamage effects and the very high acquisition speed make light-sheet fluorescence microscopy stand out.

The MuVi SPIM, the InVi SPIM, the QuVi SPIM, the LCS SPIM and the TruLive3D Imager, as well as the solutions for photomanipulation, have been built up around your sample to meet the requirements of your application for fixed or cleared samples. Take a look at the different examples.

Embryogenesis & Developmental Biology

Light-sheet fluorescence microscopy enables the observation of events in real time for several days.

The MuVi SPIM was conceived for gentle, large specimen imaging. It features four orthogonal views of the sample without the need for rotation. The low magnification, high NA objective lenses enable fast, high-resolution, large field-of-view imaging of entire embryos as well as tissue and organ development tracking.

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

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Drosophila Embryo

Drosophila embryo expressing H2A-mCherry and MS2-GFP. Imaged on the MuVi SPIM.

Courtesy of:
Jared Toettcher
Princeton University, Princeton, NJ, USA

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Drosophila Egg chambers

Drosophila ovariole stained with phalloidin to label actin (red) found along membranes and in 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. Imaged on the MuVi SPIM.

Courtesy of:
Jasmin Imran Alsous
Schvartsman Lab
Princeton University, Princeton, NJ, USA


3D Imaging of Organoids

Organoid research has been established as an essential tool for studies in cancer research, drug discovery and regenerative medicine.

The InVi SPIM singles out in the gentle handling of the most delicate samples and the minimization of required specimen medium. This is achieved by the incubation capacities and the V-shaped sample chamber. If high-throughput is also desirable, the QuVi SPIM or the TruLive3D Imager.

Pancreatic Spheres

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

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.

Courtesy of:
Yassen Abbas
Turco Lab, University of Cambridge
Cambridge, UK

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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

 

 

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3D Imaging of an 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 labelling, while the Gaussian beam was optimal for the mRFP labelling.

Courtesy of:
Martin Stöckl
University of Konstanz
Germany

Colonies of mouse embryonic stem cells

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


Live Imaging of Plants

The application of light-sheet fluorescence microscopy for plant research has allowed in vivo studies of plant cell and tissue biology.

In combination with other approaches, imaging of cell morphology, migration and organization, as well as tissue structure enable a better understanding of function and dynamics. The MuVi SPIM, but also the InVi SPIM provide the relevant tools for the research on the field.

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

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Arabidopsis Leaf

Arabidopsis leaf labelled with mCherry, 150 µm max projection, 0.5 µm step size. Imaged on the InVi SPIM Lattice Pro.

Different illumination patterns (i.e. Gaussian beam and Bessel beam) were selected to obtain high-resolution and large FOV.

Courtesy of:
Dr. Amrit Bhal
Department of Biological Sciences, National University of Singapore
Singapore

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Arabidopsis Root Growth Microalgae

 Autofluorescence in microalgae. Imaged on the InVi SPIM.

 


Cell Culture Imaging

Light-sheet microscopy applied to cell culture imaging reduces negative effects on cell behavior and development.

The InVi SPIM enables gentle handling of the most delicate samples and the use of small volumes of mounting media. The TruLive3D Imager further enables multi-sample imaging. In both cases, precise control of CO2, O2, temperature and humidity provides close-to-natural conditions to the sample. The QuVi SPIM further brings high-throughput imaging capabilities.

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 pre-implantation 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 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 Cells Culture

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 & Neurodevelopment

Light-sheet fluorescence microscopy has enabled imaging of the central nervous system including large networks of neurons, and even whole cleared brains (e.g. mouse).

The MuVi SPIM LS, the InVi SPIM and the QuVi SPIM are suited for in-vivo imaging of nervous system development in small embryos like Drosophila, Zebrafish and C. elegans, while the MuVi SPIM CS was conceived to image large cleared samples.

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

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

 

 

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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


Cleared-Sample / Cleared-Tissue Imaging

Clearing tissue techniques modify the optical properties of usually opaque samples to render them transparent while keeping their structure intact.

The MuVi SPIM CS, the LCS SPIM and the QuVi SPIM provide innovative solutions in sample mounting, sample size and optics for best-in-class 3D imaging of cleared samples.

Cleared Mouse Brain

Transgenic mouse expressing the fluorescent protein YFP in the brain. The sample was cleared with Clarity and imaged on the MuVi SPIM CS.

Courtesy of:
Dr. Zhang Dan
Tsinghua University
China

Cleared Mouse Brain Slice

Brain slice (approx. 500µm thick) cleared by Expansion technology. Imaged on the MuVi SPIM CS.

Courtesy of:
Dr. Peng Yuwei
Tsinghua University
China

Cleared Mouse Lymph Node

Cleared mouse lymph node. High endothelial venules (642 nm, red) and autofluorescence (488 nm, green) to visualize surrounding tissue. Imaged on the MuVi SPIM CS.

Courtesy of:
Jens Stein
University of Bern
Bern, Switzerland

Mouse Lymph Node

BABB-cleared mouse lymph node. High endothelial venules (red) and autofluorescence (green) to visualize surrounding tissue. Imaged on the QuVi SPIM.

Courtesy of:
Jens Stein
University of Bern
Bern, Switzerland

Mammary Gland Cleared Tissue

iDisco mammary gland cleared tissue (approx. 4mm x 13 mm x 5 mm) showing the ducts. Imaged on the MuVi SPIM CS.

Courtesy of:
Dr. Wenqian Song
Shanghai Institute of Biochemistry and Cell Biology, CAS
China

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Cleared Mouse Head

Labeled with anti-tuj1 (green) to mark developing nerves and with anti-desmin (red) to mark differentiating muscles. Tiled image (6 × 5). Imaged on the MuVi SPIM CS.

Courtesy of:
Glenda Comai
Institut Pasteur
Paris, France

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Neuronal Network

Neuronal network (stained with GFP) of a CUBIC-cleared mouse brain

Color-coded depth representation: maximal depth displayed 1.23 mm.
Imaged on the MuVi SPIM CS.

Courtesy of:
Montserrat Coll Lladó
European Molecular Biology Laboratory (EMBL)
Barcelona, Spain


Other LSFM Applications

Many other applications can benefit from the advantages of imaging with light-sheet microscopy, e.g. marine biology.

Take a look at the different systems, the MuVi SPIM, the InVi SPIM, the QuVi SPIM, the LCS SPIM or the TruLive3D Imager, and select the one that best fits your sample needs.

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

 

 

Annelid Embryo

Annelid Embryo

Platynereis dumerilii (annelid) in embryonic stage. Stained for EMTB, a microtubule associated protein. Color-coded depth representation. Imaged on the MuVi SPIM.

Courtesy of:
Hella Hartmann, Mette Handberg-Thorsager, Pavel Tomancak and Yu-Wen Hsieh
TU Dresden and Max Planck Institute of Molecular Cell Biology and Genetics (MPI-CBG)
Dresden, Germany