In recent years, 3D cell cultures have gained popularity due to their clear potential to mimic a realistic in vivo tissue in terms of structure, architecture, multi-cellular microenvironments and function. Established 3D cell culture systems such as organoids and spheroids have been proven to be valuable tools in applications including disease modeling, cancer research (e.g. tumor biology and host-tumor interactions), drug development and testing, as well as regenerative medicine.
Light-sheet microscopes are the most powerful tools for investigating the complex structure and behavior of 3D cell cultures by performing live, fast, long-term and high-resolution imaging with minimal phototoxic effects.
Bruker offers different light-sheet geometries to enable multiplexing and gentle, long-term 3D cell culture imaging under precisely controlled environmental conditions.
hESC-derived pancreatic spheres. Imaging on the TruLive3D Imager enables collecting information of several samples in one experiment. Visualization: Imaris (Bitplane)
Yung Hae Kim
Graphin-Botton Group, MPI-CBG
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
Spheroid stained with anti-GFAP (Alexa 488) to label astrocytes and anti-Neurofilament200 (Alexa555) to label neurons. Imaged on the InVi SPIM.
AG Leist, University of Konstanz
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.
University of Konstanz
Colonies of mouse embryonic stem cells stably expressing H2B-mCherry and IRFP670 with a membrane-targeting signal. Imaged on the InVi SPIM.
European Molecular Biology Laboratory (EMBL)
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).
European Molecualr Biology Laboratory (EMBL)
Sample: HeLa cells (Neumann et al., Nature. 2010 Apr 1;464(7289):721-7)
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 cell line (BS-C-1) stained for f-actin and chromatin. Image taken with InVi SPIM Lattice Pro.
Nikon Imaging Center
University of Heidelberg