Platform A: Techniques and Technology Development
| Platform Coordinators | |
|---|---|
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Institute for Cardiovascular Physiology Goethe-University Frankfurt Theodor-Stern-Kai 7 60590 Frankfurt/Main |
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Pulmonary Pharmacotherapy Justus-Liebig-University Giessen Aulweg 130 35392 Giessen |
The ECCPS provides a broad spectrum of technologies for the analysis of the cardio-pulmonary system, ranging from techniques to study single molecules through pre-clinical phenotyping models to human-related technology e.g. biomarkers, biobanks and interventional trials.
Planned service units for the second funding period:
- Bioinformatics and deep sequencing (service oriented bioinformatics facility, deep sequencing analysis platform, integration of "omics" technologies)
- Cardio-pulmonary deep phenotyping (stress test and mouse disease facility, small animal imaging platform)
- Molecular analysis and high resolution imaging (molecular in vivo imaging, molecular analytic platform)
Technologies currently available in the ECCPS
| Level | Techniques |
| Genomics, Transcriptomics | cDNA microarray platform including bioinformatics, chip manufacturing, DNA sequencing,SNPs, STRs, chromatin immunoprecipitation (ChIP), two/three-hybrid screens, microarraysfor protein-DNA interaction, RNA fingerprinting /RAP-PCR, SNP pyrosequencing laser-mediatedmicro dissection, DNA methylation assay |
| Proteomics | MALDI-TOF/TOF-MS, ESI-MS/MS, SELDI-MS, Nano-LC-ESI-MS/MS, protein profiling onantibody chip, DNA-assisted protein chip, ELISPOT, micro 2-D protein-analysis, tissuemicroassay, peptide mapping & sequencing, cross-linking analysis, phosphoproteome, phage-display screen, surface plasmon resonance |
| Lipidomics, Glyconomics | HPTLC, ESI-MS, LC-MS/MS, 2D-HPLC, GC/MS, MALDI-TOF/TOF-MS, nano-ESI-MS/MS, carbohydrate composition & linkage analysis, LC-MS/MS |
| Protein Biochemistry | Isothermal titration calorimetry, fluorescence spectroscopy, fluorescence resonance energy transfer between site-specific fluorescently labelled macromolecules, circular dichroism, cross linking of site-specific chemically modified macromolecules, fluorescence stopped flow, quenched flow, SPOT analysis, chromatography units |
| ROS/NO Detection | ESR spectroscopy, in-situ remission spectroscopy, chemiluminescence, single cell microfluorometry, respiratory measurements |
| Imaging - Molecular | FRET, BRET, FRAP, confocal microscopy, electron microscopy, automated high throughput in situ hybridization, in-situ PCR, immunocyto(histo)chemistry, cell-edge-detection system for determination of contractile function, cell coupling; fluorimetric second messenger detection, activatable smart probes, live cell imaging |
| Imaging - Small animal | Small animal NMR, micro-CT, flat-panel volumetric CT, mouse echocardiography, hybrid fluorescence molecular tomography-CT |
| Imaging - Morphometry | Software-assisted vascular morphometry (medial wall thickness, lumen area, degree of muscularization), alveolar morphometric routines (septal wall thickness, mean alveolar intersept & airspace), heart stereology, assessment of innervation |
| Imaging - Clinical | Cardiac MRT, IVUS, MS CT, virtual imaging, 3-D echocardiography |
| Cytometric Profiling | FACS-based multiparametric immunophenotyping/sorting , high volume immunomagnetic cell sorting (autoMACS); analysis of cell-specific gene & protein expression (multicolor fluorescent antibodies) in complex cell samples, ELISPOT |
| Transfection Technology | Adeno-, retro-virus & bacteria mediated gene transfer, liposome based gene transfer technologies, PEI-DNA aerosol technology, aerosolized nanocarriers for gene transfer, ribozymes, antisense technology, VP22-mediated protein-transfer |
| Gene Silencing | siRNA manufacturing, siRNA aerosolization & vector technology, conditional RNAi in mice (Tet control, Cre/lox control), lentiviral siRNA transfer |
| Stem Cell Technologies | Large scale murine ES-cell production induced ES-cell differentiation and targeted manipulation of cell development. Purification of human & murine progenitors, circulating cells & (cancer-) stem cells. In vivo tracing of stem cell progeny with reporter-transgenes, TK constructs, & radioactive labelling, GMP facilities & permission for the isolation & cultivation of human stem cells for clinical use (production license HA-FE-504/A) |
| Angiogenesis Models | In-vitro assays (Fibrin & Matrigel assays), Matrigel plug assays in mice, hind limb ischemia model, tumour angiogenesis models, ES cell angiogenesis assay |
| Transgenic Mouse Facility | ES cell bank, generation of transgenic mice, knock-out/ -in mutants, of cell-type-specific temporally-controlled somatic mutants, direct links to several mouse consortia (IKMC, EUCOMM, GGTC, DiGtoP, NGFNplus, EMMA, NGFNplus) |
| Mouse Phenotyping | Invasive, non-invasive & telemetric hemodynamic assessment, echocardiography, mouse lung & airway (bronchoconstriction, mucociliary clearance) function testing, precision cut lung slices |
| Mouse Models of Disease | Cardiac ischemia & infarction, left & right heart hypertrophy & failure, atherosclerosis, restenosis, allograft vasculopathy, pulmonary hypertension, lung fibrosis, pulmonary emphysema / COPD, ARDS, ventilator-induced lung injury, bacterial and viral pneumonia, emphysema, sleep apnoea, lung tumour & metastasis, thrombosis models |
| Zebrafish & C. elegans | Breeding, transgenic fish, models of cardiac & vascular development, manipulation, generation & characterization of C. elegans |