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Techniques and Instrumentation

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cell culture lab Geesthacht

The infrastructure reflects the interdiciplinary character of the department and combines the requirements of biological and material research. The laboratories in Geesthacht are classified for working under safety level 2 (S2) according to Biological Safety and S1 for reseach with genetically modified organsism (GMO).

Several different primary and secundary cell types are used as targets for analysing cell-materíal interactions. Preferably human mesenchymal stem cells are cultured and differentiated into the desired cell line of the musculco-sceletal system.
In order to test the antibacterial properties of selected materials and material surface treatments a set of gram-negative and gram-positive bacteria are available.

Selected components of the infrastructure

  • Cell culture safety cabinets and incubators
  • Cell counting and viablity by Roche Casy®cell counter and analyser
  • Bioreactor(s) and sensor
  • Flow cytometer S3e cell sorter (BioRad)

The cell response to material contact occurs at different level of biological organisation. Therefore, gene expression analyses are combined with proteomic studies and functional cell assays.

Selected components of the infrastructure

  • PCR (Eppendorf Mastercycler)
  • Realtime-PCR (BioRad C1000 Thermo Cycler CFX96)
  • Fluorescence or luminescence measurements (PerkinElmer Multiplate-Reader Victor 3-V, Hitachi F 2500 Fluorescence Spectrophotometer)
  • NanoDrop (2000c Spectrophotometer)
  • Imaging of agarose- and polyacrylamide gels (BioRad ChemiDoc MP)
  • Blotting system (Life Technologies iBlot)
  • Multi-parameter analysis of selected proteins as indicators for cell responses by BioPlex (BioRad)

The fundamental assessment of cell responses has to be done by microscopic observation.

Selected components of the infrastructure

  • Fluorescence microscopes (e.g. Nikon TI Eclipse)
  • Laser Scanning Confocal Microscopy (Leica TSC SP8)
  • Scanning Electron Microscope
Fittosize _780 0 81474ded5b367095fbaf4fd2f1bee0ae Sample Mounting Fw Jm

Hereon operates several synchrotron beam lines for engineering materials science at the PETRA III source of DESY in Hamburg. Among them an imaging beamline (IBL) P05 dedicated to micro- and nanotomography at energies between 5 and 50 keV and a scanning small/wide angle X‐ray scattering (SAXS/WAXS) nanofocus end station at P03.

Our specialty is the development of sophisticated sample environments optimised for the investigation of metallic biomaterials at these beamlines.

The objective is to gain information with highest spatial and density resolution from explants about structure and composition of bone‐implant interface. Beside micro‐ and nanotomography also scanning SAXS is available with an ultra‐small beam to obtain information about the bone structure formed around the implant.

Selected components of the infrastructure

  • The microtomography end station at IBL is equipped with sample changing robot and several sample environments (furnace, cooling cell, corrosion cell, load frame, etc). Imaging techniques available: • Absorption contrast with high density and spatial resolution
    • Inline (propagation-based) phase contrast for soft/biological tissues
    • Differential phase contrast (DPC) using a grating interferometer for objects with mixed phase and absorption contrast
  • Electron‐induced X‐ray emission spectroscopy (EIXE) combined with SRμCT
  • Scanning SAXS/WAXS with a 250 x 350 nm beam size (nanofocus end station)
  • Laboratory SAXS/WAXS setup (Bruker Nanostar) and 2D/3D X-ray analysis (µCT, YXLON Y Cougar) for pre-studies

Medical imaging is essential for the development of novel implants, especially if they actively intervene in the metabolism of the body.
Therefore, the Division Metallic Biomaterials (WB) operates an outstation at the Molecular Imaging North Competence Center (MOIN CC) in Kiel as a platform for multi-modal imaging in various animal models.
The goal is to bring together MR method development and metallic implants, to expand our in vivo activities on materials developed at WB, and to make a significant contribution to the development of novel implants in the long term.

For this purpose, longitudinal studies are carried out to provide high-resolution temporal and spatial information on physiological changes, the degradation behavior of the material, the rate of bone formation and its structure and quality.
In addition, we perform cell biology and histology on explants and use immunohistochemistry for a more detailed analysis.

Our specialties include the evaluation of bone healing processes in response to Mg implants in wild-type and diabetic animal models, the assessment of angiogenesis with magnesium-based implants, and the evaluation of the impact of magnesium alloys on macrophage polarization by multimodal imaging.

Selected infrastructure components

  • Optical imaging Whole body in vivo epi-fluorescent and bioluminescent imaging
    (NightOWL II, Berthold Technologies).
    In vivo Fluorescent tomography
    (FMT 2500, PerkinElmer)
    Endoscopic confocal fluorescence imaging
    (Cellvizio, Mauna Kea)
    Intra-operative fluorescence microscopy
    (modified surgical microscope, Möller-Wedel, with technical adaptations developed by MOIN CC)
    Multi-photon microscopy
    (TriMScope, LaVison)
  • Micro-computed tomography (VivaCT 80 micro-CT, SCANCO)
  • Magnetic Resonance Imaging (MRI) and Spectroscopy (MRS) (7 Tesla ClinScanTM, Bruker BioSpin)
  • Ultrasound (US) and Photoacoustic Imaging (USPA) (Vevo 770™ ultrasound system and Vevo® LAZR, Visual Sonics)
Molecular Imaging North Competence Center (MOIN CC)