14. April 2021
Immunophenotyping of Tissues Using Digital Techniques
What is Immunophenotyping?
Immunophenotyping refers to the identification of specific phenotypic cells using markers or antigens which are present on the cell’s surface, cytoplasm, or within the nucleus. It aids the classification of cell lineage using antibodies targeting specific markers or antigens on the cells.
Immunophenotyping can be used to diagnose various types of diseases including cancer by comparing normal cells with cancer cells in suspension (i.e. leukemias) or tissues (i.e. solid cancers).
Types of Immunophenotyping
In immunophenotyping, two types of tests are used: flow cytometry for fluid suspension samples and immunohistochemistry for cells as well as tissue sections on slides. Flow cytometry is used to analyze single cells in suspension including bone marrow, peripheral blood samples or dissociated tissues. It is a fast and reliable method of immunophenotyping which can be used to detect infrequent hematological malignancies, clonality in B-cell lymphomas and other disease-specific cellular phenotypes with ease. Flow cytometry can measure cell size, cell granularity, total DNA, surface and intracellular proteins and therefore is an extremely powerful method for immunophenotyping. The only disadvantage is that cells are analyzed in suspension and valuable information in respect to their native tissue context is lost.
Immunohistochemistry involves the staining of tissue sections, smears, TMAs, or cultured cells on slides. Immunophenotyping can be performed with tissue cytometers, the workflow includes the whole slide scanning of tissue sections and the follow up analysis of single cellular phenotypes in tissue context.
Immunophenotyping solutions from TissueGnostics
The TissueFAXS® platform is a next-generation tissue cytometer. It couples the advantages of innovative high-resolution IF multi-channel microscopy and automated whole slide imaging with the analytical accuracy of flow cytometry immunophenotyping. The big advantage of this is that tissue context is still available while whole slide images – digitized slides – can be easily shared and discussed with colleagues via telepathology.
The TissueFAXS® tissue cytometer is successfully employed in various research fields including cancer, pathology, developmental biology, immunology, urology, dermatology, and drug development. It satisfies the demands of each of these disparate analytical fields through its extreme modularity. Although based on the same platform, TissueFAXS® cytometers feature integrated solutions that enable unique functionality for critical aspects of study. The TissueFAXS Q comes with a high-tech spinning disk unit for widefield or confocal imaging using a single physical disk. The TissueFAXS PLUS, meanwhile, is equipped with an 8-slide stage used to scan samples in both brightfield and fluorescence modes. Each of these platforms is modular and upgradeable to meet the changing demands of your research area.
At TissueGnostics, our analysis algorithms typically begin with single cell assessment which requires a master channel, usually DAPI fluorescent stain. Nuclei are then segmented using a contour overlay which enables the measurement of relative staining intensity for markers both inside and outside the nucleus. Using these measurements as our baseline we can also identify multicellular objects in tissue for detecting more precise, tissue contextual data.
If you would like to learn more about immunophenotyping solutions from TissueGnostics, contact us today.
Are you also interested in A brief Introduction to Automated Tissue Classification? Click here