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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/FCCS/iFCS), as well as combined with superresolution for nanoscale dynamical studies (STED-FCS). Moreover, light-sheet fluorescence microscopy support allow users to image live organisms and/or optically cleared larger samples at unprecedented volumetric speed with low phototoxity. The newly installed lattice light-sheet microscope furthermore allows fast high-resolution whole cellular dynamical imaging.

The Advanced Light Microscopy facility is further connected to research environments developing RESOLFT microscopy (Testa-lab, http://www.testalab.org) as well as nanoscale molecular mechanism tools (Sezgin-lab, https://www.csi-nano.org).

 

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SERVICES

Service 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, including fast cellular dynamics

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/FCCS/iFCS, STED-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 and membranes

Light-sheet/Lattice 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)
  • Fast high-resolution volumetric cellular imaging (LLSM)

EQUIPMENT

  • PALM/STORM. Zeiss Elyra 3D PALM.
  • STED. Leica SP8 3X STED with FALCON FLIM/FCS
  • SIM. Zeiss Elyra 3D SIM
  • FCS. Zeiss LSM780
  • Light-sheet. Zeiss Z.1
  • Lattice light-sheet. Zeiss 7

RECENT PROJECTS

SIM, STED, STORM/PALM

  • Correlative STED super-resolution light and electron microscopy on resin sections, J.Phys.D: Appl. Phys. (2019)
  • Mic60 exhibits a coordinated clustered distribution along and across yeast and mammalian mitochondria, PNAS (2019)

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 

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