Techniques used by the Radiotherapy & OncoImmunology Laboratory
The biology of tumors is studied at the macroscopic (PET) and microscopic (cell, subcellular) level.The aim is to compare different functional imaging modalities for the same tumor.
The focus is on:
Quantitative immunohistochemistry:
The backbone of this system are:
- the vascular architecture (9F1 or CD31/24)
- tumor blood perfusion (Hoechst 33342)
- exogeneous markers/indicators of hypoxia (CA-IX, pimonidazole)
This system can be extended by:
- proliferation (BrdUrd, IdUrd, Ki67)
- growth factors ((p)EGFR, (p)AKT)
- DNA damage (gamma-H2AX, 53BP1)
At the microscopic level, the tumor biology is studied in a quantitative manner with preservation of the tissue architecture and spatial associations.
Therefore, we have developed a system for co-registration and quantitative analysis of microenvironmental phenotypic tumor charateristics. The method is based on immunohistochemical detection of multiple fluorescent signals in complete tissue sections.
Staining:
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Immunofluorescence (DAPI, Fitc, Tritc, Far Red) |
Immunohistochemistry (DAB, Hematoxyline) |
Quantification:
Components of our digital imaging systems:
- multi-color fluorescence microscopy
- computer-controlled scanning and imaging system:
- CCD camera
- motorised scanning stage
- image acquisition and analysis software
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| System |
Scanning yields a composite image for a marker |
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| Multiple color image after combining the sequential recorded composite images |
| Our laboratory has permission for genetically modified organisms ('ML-II'). The facilities are:
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| Hypoxystation |