The Immunohistochemistry laboratory operates in the field of translational research dealing with the clinical validation of protein and molecular markers with analytical methods such as immunohistochemistry, immunocytochemistry, immunofluorescence, fluorescent and chromogenic 'in situ' hybridization, Tissue Microarray, digital slides and image analysis with artificial intelligence algorithms.
In the Immunohistochemistry laboratory, investigations are performed on histological, cytological and cryostatic material for research, diagnostic and predictive purposes of response to targeted therapies (target therapy). The main objective of the research activity is to deepen the knowledge of molecular mechanisms involved in cancer diseases using differentiation biomarkers and cellular function. In particular, projects evaluate how different markers are expressed in different tumor histotypes and how these may influence tumor progression, as well as have prognostic/predictive and therapeutic value. The laboratory is equipped with a number of highly innovative instrumentation including a Tissue Microarray creation instrument (TMA Master-3DHYSTEC) in order to simultaneously evaluate large sample series with significant reagent savings and optimization of standardization of the investigations performed. The Ventana DP 200 slide scanner (Roche) allows obtaining bright-field digital slides of 6 preparations simultaneously that can be used for image analysis with artificial intelligence algorithms (QuPath software). Image analysis allows quantitative evaluation of a large number of morphological and morphometric parameters (such as number of tumor cells, tumor area and perimeter, H-score value) useful to validate the research data obtained from 'light microscope observation. Finally, the daily use in diagnostic activities of three BenchMark Ultra (Roche) immunostainers for immunohistochemical analysis, in situ hybridization analysis, and SISH analysis allows for the constant standardization of methods and biomarkers often used in the research setting.
All investigations performed in the laboratory take into account specific protocols defined at the national level by the scientific societies of medical oncologists (AIOM) and anatomic pathologists (SIAPEC-IAP). The activities of the laboratory have been validated over time through participation in the Italian CQ RER-Bologna quality control and European quality control programs NordiQC.
Main lines of research
The main objective of the research activity is translational and consists mainly in deepening the knowledge of the molecular mechanisms involved in oncological pathologies. In particular, the analyses are aimed at obtaining information on the immunophenotype of cells and tissues by means of recognition implemented by specific antibodies or probes, making use of immunological and hybridization reactions, in order to define the cellular composition of disease processes, identify pathogens, and detect biomarkers useful for diagnosis, prognosis, and prediction of therapeutic response of human cancers.
The most impactful applications involve early diagnosis of malignancy, identification of molecular mechanisms of malignancies, and research and clinical validation of markers predictive of prognosis and response to targeted biological therapies (target therapy). The recent introduction of instrumentations such as Tissue Microarray (TMA Master-3DHYSTEC) and Ventana DP 200 allow the analysis of large, even retrospective case series and enable a type of translational research that bridges genetics, pathology and clinical analysis. The laboratory constantly collaborates with various study groups both of our University and University Hospital Company and other centers and is the main reference center for these applications within the University.
The Laboratory offers the possibility of internships and traineeships to students, residents of the University and support to doctoral students engaged in medical research.
Techniques in use
- Immunohistochemistry and Immunocytochemistry
- Immunofluorescence
- Hybridization in Situ
- SISH
- Tissue Microarray (TMA)
- Digital slide production
- Imaging analysis.
Main equipment provided
N. INVENTORY | SPECIFIC NAME OF INSTRUMENTATION | CONTACTOR | UBICATION | HOME USE OF INSTRUMENTATION | TIPOLOGY OF USE | TEMPES OF USE |
| 55/001 | TMA Master | 3Dhistech | UO Pathology Anatomy-First Floor Room 012 | Tissue Microarray | Creation kerosene blocks of tissue microarrays according to specific layouts, on which carried out different laboratory methods (IHC, ISH) | variable |
| 661749 | Ventana DP200 | Roche | UO Pathology Anatomy-First Floor Room 012 | Scanning slides in bright field (20X or 40X) for image analysis | Tool used to quickly scan all types of slides on which to perform digitized analysis with specific programs (example Qupath) | weekly |
| 623265 | BenchMark Ultra | Roche | UO Anatomy Pathology-First Floor Room 012 | Automatic immunostainer | Instrument used for immunohistochemical analysis of primary antibodies highlighted by different detection systems , in situ hybridization and immunofluorescence analysis | daily |
| 623266 | BenchMark Ultra | Roche | Pathological AnatomyUO-First Floor Room 012 | Automatic Immunocolouriser | Instrument used for immunohistochemical analysis of primary antibodies highlighted by different detection systems , in situ hybridization and immunofluorescence analysis | quotid |
| 623267 | BenchMark Ultra | Roche | Pathology Anatomy Unit-First Floor Room 012 | Automatic immunostainer | Instrument used for immunohistochemical analysis of primary antibodies highlighted with different detection systems , in situ hybridization and immunoflorescence analysis and SISH (two-color chromogenic in situ hybridization) | daily |
