Advanced Light Microscopy & Image Analysis — Technology Platform offers several high end state-of-art microscope setups for a wide range of imaging techniques and applications.

Many of the setups are equipped for live imaging.

Confocal and Two-Photon Microscopy 

Our confocal microscopes are equipped with multiple laser lines, various objectives and spectral detector units (using photomultiplier tubes – PMTs – or high sensitivity detectors). Two-photon excitation is possible for thick samples. Observation of subcellular structures can be done with the high-resolution Airy scan detector. A dedicated FLIM microscope can be used to study intracellular protein-protein interactions.

More information is available for internal users here.

Instruments

• Leica Stellaris 8
• Leica TCS SP8
• Leica TCS SP8 with Digital LightSheet option
• Zeiss LSM 700
• Zeiss LSM 880 NLO (with two-photon excitation)
• Zeiss LSM 980 Airyscan
• Nikon/​Andor CSU‑W1 (spinning disk)
• Becker & Hickl FLIM (fluorescence lifetime imaging)

Some Applications

• Sequential multi-color fluorescence imaging (UV/green/red/far-red dyes) of adherent fixed cells and tissue sections up to 100 µm thickness
• z‑stacks for 3D reconstructions
• Tile scans for large area samples
• Multiposition imaging
• Spectral unmixing of overlapping emission spectra (also using white light lasers)
• Confocal imaging with Airyscan technology (about two-fold improved resolution, speed and sensitivity)
• Long-term live imaging of adherent cells
• Live cell imaging with very fast time series (e.g. vesicle movement, Ca²⁺responses)
• FRET (Förster resonant energy transfer) for detecting protein-protein interactions in situ measured via sensitized emission (fluorescence intensity ratio) or fluorescence lifetime imaging (FLIM)
• Laser photomanipulations such as FRAP (fluorescence recovery after photobleaching) or photoswitching/​photoactivation
• 2‑Photon excitation for imaging in thick samples
• (Intravital) imaging of small organs/​organisms with long working distance objective and/​or 2‑Photon excitation and non-descanned detection
• DIC microscopy
• Combination of confocal and selective plane imaging on the same system with the Digital LightSheet technology

Two-Photon Microscopy for in vivo deep tissue imaging

Deep tissue and organ imaging is possible due to the high penetration depth of the infrared radiation in Two-Photon microscopy. Our dedicated setup is completed with equipment for maintaining conditions for long-term live imaging. Large area imaging is possible for cleared or un-cleared tissue samples.

More information is available for internal users here.

Instrument

LaVision Biotec Trimscope II

Some Applications

• Two-photon fluorescence imaging for non-transparent tissues labelled with fluorescent proteins, selected fluorophores or quantum dots using up to three different excitation wavelengths and six detection channels simultaneously and possibilities for spectral unmixing
• z‑stacks, tile scans and multipostition imaging
• in-vivo tracking of cells expressing fluorescent proteins
• Imaging of blood vessels labelled with fluorescent dextranes or quantum dots
• SHG (second harmonic generation) of collagen fibres
• Functional imaging of intracellular signals such as Ca²⁺ activation
• FRET imaging of endogenously expressed biosensors via sensitized emission (fluorescence intensity ratio) or FLIM (fluorescence lifetime imaging)
• Label free imaging of cellular autofluorescence (NADH, FAD)
• Photoactivation of bacteriorhodpsins for optogenetic studies

Light sheet microscopy

Our light sheet microscopes are applied for fast imaging of large volume tissue and organ samples after clearing. We provide solutions for active clearing and labelling. We also offer conditions for long-term imaging of live small transparent samples.

More information is available for internal users here.

Instruments

• Zeiss Lightsheet 7
• Miltenyi UltraMicroscope Blaze^TM
• LaVision Biotec Ultramicroscope I
• LifeCanvas Tissue Clearing and Labeling Device

Some Applications

• Selective plane illumination for 3 dimensional imaging of large samples
• Multiview acquisition with different rotation angles
• Multicolor fluorescence imaging (blue/green/red/far-red dyes) of cleared samples like spheroids, tissue and organs, or transparent specimen like Zebrafish embryos or Drosophila larvae
• Long-term and short-term live imaging

Superresolution & TIRF Microscopy

Breaking the resolution limit is possible using several microscopy techniques. We offer wide field TIRF microscopy for studying dynamic membrane processes in live cells and possible applications to localisation microscopy (e.g. PALM). Our STED microscope can be used for both fixed and live samples as well as for the RESOLFT technique.

More information is available for internal users here.

Instruments

• Olympus TIRF
• Nikon TIRF (with 4 EMCCD cameras)
• Abberior STED

Some TIRF applications

• observation of membrane-associated cellular processes
• FRET microscopy by sensitized emission
• fast, sensitive single molecule imaging and tracking

Some STED applications

• subdiffraction-resolution imaging of subcellular structures and organelles in membranes, cytoplasm and nucleus of flat adherend cells and thin tissue sections
• multicolor superresolution imaging

Wide field microscopy & laser microdissection

Laser microdissection is applied for specific isolation of individual cells or larger areas of a tissue sample, which will undergo further molecular analysis, i.e. contamination-free molecular analysis of DNA, RNA and proteins.

More information is available for internal users here.

Instrument

• Olympus CellSens
• Leica wide-field system
• Zeiss PALM MicroBeam (Axio Observer Z1)

Some applications

• Multicolor wide-field fluorescence imaging of adherent cells and thin tissue sections (up to 10 µm thickness)
• Tile scan and multiposition imaging for large area samples on 1 – 4 microscope slides
• Multi-well plates imaging
• Long-term imaging of live cells
• Imaging of fast intracellular processes (e.g. Ca2+ imaging)
• Imaging of biosensors and of protein-protein interactions using FRET (Förster resonant energy transfer) via sensitized emission (fluorescence intensity ratio)
• Prefinding of cells and regions using multicolor wide-field fluorescence imaging (software: ZEN Blue), followed by laser-guided precise microdissection (software: PALM Robo) of histological sections (up to 100 µm thickness) on glass or membranes slides and of living cells in membrane dishes

Ultra-high content microscopy 

The ultra-high content microscopy system on our platform is based on cyclic immunofluorescence imaging technology, which allows for sequential imaging of tens to hundreds of different cellular markers. A fully automated workflow is set up to perform labeling, imaging in 4 spectral channels, and removal of the fluorescent signal (via photobleaching) or the labeled antibodies (via enzymatic reaction). The omics datasets are analyzed with dedicated software to generate complex interpretations of the spatial biology data.
 

Instrument:

• Miltenyi MACSima platform
   

Some applications:

• Ultra-high-plex spatial protein profiling with panels featuring over 200 validated protein markers
• High-plex imaging of 27 RNAs
• Combined protein and RNA detection within the same tissue section
• Applicable for both mouse and human tissues and cell lines (with PFA or FFPE fixation methods)