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Advanced Light Microscopy

National facility

The mission of the Advanced Light Microscopy facility is to give support with advanced fluorescence microscopy for nanoscale biological visualization using SIM, STED, STORM/PALM superresolution imaging. In addition the facility support single molecule spectroscopy measurement and analysis with fluorescence correlation spectroscopy (FCS). Moreover, light-sheet fluorescence microscopy support allow users to image live and/or optically cleared larger samples at unprecedented volumetric speed with low phototoxity.

SERVICES

Support is done to all stages of the project including (pre)planning, sample optimization, fluorescent probe selection, image acquisition, and post-acquisition image processing.

  • Super-resolution microscopy (SIM, STED, STORM/PALM) - nanoscale cellular imaging of fixed or living samples, and cleared/expanded tissue
  • Fluorescence Correlation Spectroscopy - single molecule spectroscopy measurement and analysis to evaluate interaction, aggregation, mobility, dynamics
  • Light-sheet microscopy -fast optical sectioning of large volumes of organoids, organisms, cleared/expanded tissues

Collaborative project management and transfer of unique knowledge to individual researchers are supported nationally, including organization of workshops and courses in advanced fluorescence imaging and spectroscopy applications and method developments.

APPLICATIONS

SIM, STED, STORM/PALM

  • Investigation of molecular architecture of sub-cellular entities.
  • Organization of protein localization in cell biology with sub-diffraction resolution.

FCS

  • Analysis of receptor-receptor, ligand-receptor, protein-protein, protein-peptide and protein-vesicle interactions in solutions and in living cells
  • Aggregation and affinity analysis at single molecule concentration levels in solutions

Light-sheet

  • Morphogenesis, Embryogenesis and Organogenesis (e.g. in Zebrafish)
  • 3D cell culture imaging using spheroids, tissue and organotypic culture
  • Imaging optically cleared/expanded samples (e.g. CLARITY, DISCO, ExM)

EQUIPMENT

  • PALM/STORM. Zeiss Elyra 3D PALM.
  • STED. Leica SP8 3X STED and Leica SP5 NIR-STED
  • SIM. Zeiss Elyra 3D SIM
  • FCS. Zeiss LSM780
  • Light-sheet. Zeiss Z.1

RECENT PROJECTS

SIM, STED, STORM/PALM

  • Sexual dimorphism in the width of the mouse synaptonemal complex, Journal of Cell Science (2018)
  • Extracellular nanovesicles released from the commensal yeast Malassezia sympodialis are enriched in allergens and interact with cells in human skin, Science Report (2018)
  • Neuronal Aβ42 is enriched in small vesicles at the presynaptic side of synapses, Life Science Alliance (2018)
  • Active and inactive β1 integrins segregate into distinct nanoclusters in focal adhesions, Journal of Cell Biology (2018)

FCS

  • Educated natural killer cells show dynamic movement of the activating receptor NKp46 and confinement of the inhibitory receptor Ly49A, Science Signaling (2018)
  • Stabilized Cyclic Peptides as Scavengers of Autoantibodies: Neutralization of Anticitrullinated Protein/Peptide Antibodies in Rheumatoid Arthritis, Chemical Biology (2018)
  • A Protein-Based Encapsulation System with Calcium-Controlled Cargo Loading and Detachment, Angewandte Chemie International Edition (2018)

Light-sheet microscopy

  • An orthotopic glioblastoma animal model suitable for high-throughput screenings, Neuro-Oncology (2018)
  • OLMα2 Cells Bidirectionally Modulate Learning, Neuron (2018)