Immunophenotyping

Immunophenotyping is a most important and regularly used technique in Immunology and Immunological research studies. The technique is usually used for the disease monitoring and diagnostic purpose either by immunohistochemistry or by utilizing the high-throughput technique Flow Cytometry.. Flow Cytometry is a common method for the immuno phenotyping the different cell subsets or sub populations of cells.

Immunophenotyping is used for the identification and characterisation of the cell surface markers and intracellular markers. In simple terms, the technique is used to detect and study the proteins expressed by the cells. Samples used for the study could be Tissue Section (Fresh or Fixed) or Cell suspension (mono-dispersed cell suspension for FCM). This technique helps in rapid and easy phenotyping of each cell in a heterogeneous sample according to the presence or absence of a protein combination. Markers (or proteins of interest) are usually functional membrane proteins which are involved in cell communication or signalling, cell adhesion or metabolism.

Fluorescent-conjugated antibodies are commonly used for the detection of the surface or intracellular proteins. (Fluorescent molecule is conjugated to an antibody, the antibody which is specific to a specific antigen present on the cells). Usually CD markers (Cluster of differentiation) are stained and investigated.

Main applications of this technique are, The detection and measurement of specific immune cell populations and subpopulations, Comparing the cancer cells to normal cells of the immune system.

Used for:

1) Lineage Assignment

2) Maturational characterization of malignant cells

3) Detection of clonality

4) Heterogeneity and aberrant features of the malignant cells

5) Quantification of Haematopoietic cells.

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