flowcytometry &cytolytics (8)

Flow Cyotmetric Immunophenotyping

Immunophenotyping enables the classification of cells beyond purely morphological assessment using the identification of cell membrane and intracellular antigens and is thus a central component of hematological diagnostics. In this examination, leukocytes are stained and sorted using immunological markers. The fluorescent dyes used allow differentiated and precise differentiation of individual leukocytes according to function and maturation stag.

What is immunophenotyping?

 

Cells of the hematopoietic system can be characterized by the detection of surface proteins. These surface proteins are usually specified according to the CD (cluster of differentiation) nomenclature. The detection of surface proteins is performed by specific antibodies coupled to a fluorescent dye. The detection of surface proteins is performed by specific antibodies. This method, which allows the measurement of antigens on a large number of blood cells in a very short time, is called flow cytometry. If, for example, lymphocytic populations are difficult to distinguish from one another under the microscope, flow cytometry can be used to quantify the ratio and number of immune cells or other cell populations of the blood or bone marrow. Immunophysiologically, different tasks can be assigned to the lymphocyte populations. Last but not least, importantimmunohistologicall findings in humans have been obtained by correlating the clinical phenotype (tendency to infection, pathogen spectrum) with the absence of certain cell populations in patients.

What is meant by Cluster of Differentiation (CD)?

 

CD molecules are membrane-bound glycoproteins, some of which are expressed in a cell-specific manner and can have a wide variety of functions: Some CDs have receptor or signaling functions, while others have been shown to have enzymatic activity. Also, some cluster molecules are thought to play a central role in intercellular communication. To date, several hundred molecules have been characterized, and it can be assumed that many more CDs exist.

What is the flow cytometry procedure for immunophenotyping?

 

The test material is usually peripheral blood and/or bone marrow, but other fluids e.g. cerebrospinal fluid or pleural effusion can also be used for testing. Based on the antigen profile of the analyzed cells, the lineage affiliation (myeloid vs. lymphoid) and the degree of differentiation can be determined. Modern multiparametric flow cytometry is based largely on advances in three areas: Laser optics, computerized data processing, and the development of new fluorescent dyes for coupling to the corresponding monoclonal antibodies. Membrane glycoproteins are detected by flow cytometry with fluorescently labeled monoclonal antibodies, usually using a combination of three or four different fluorochrome-labeled antibodies to characterize the cells. When several fluorescent dye-labeled antibodies are combined and the different scattering light properties of cells are exploited, it is possible to classify malignant hematologic neoplasms and, if necessary, to assess the success of therapy in the context of follow-up and minimal residual disease (MRD) control.

Membrane glycoproteins are detected by flow cytometry with fluorescently labeled monoclonal antibodies, usually using a combination of three or four different fluorochrome-labeled antibodies to characterize the cells. When several fluorescent dye-labeled antibodies are combined and the different scattering light properties of cells are exploited, it is possible to classify malignant hematologic neoplasms and, if necessary, to assess the success of therapy in the context of follow-up and minimal residual disease (MRD) control.

What innovation does Cytolytics offer?

 

Cytolytics offers sequential gating for detailed analysis, a graphical user interface for different computers, and a plausibly explained and understandable software language, as well as full automation of the analysis in flow cytometry. With the help of the innovation of full automation, not only large data sets are compared, but also abnormalities, anomalies, and outliers are detected based on the standardized setting. The automated analysis provides a standardized evaluation option so that familiarization with the Cytolytics software is effortless. The automated result documentation can be exported to various applications such as PPT, Word, and Excel, and a result presentation is possible directly and quickly without reprocessing. Using full automation, not only fast evaluations but also comparable, valid, repeatable, and meaningful results are delivered. With today’s technology, FACTS can identify which tumor and which cells are affected in the case of a cancer diagnosis. However, due to the time-consuming nature of gating, therapy planning and corresponding therapy control examinations often take place under great time pressure. Time is a life-saving factor and can, in sufficient quantities, enable precise, targeted therapy planning, for example in the case of leukemia. Cytolytics provides an innovative and intelligent solution for this. With time-saving analysis without gating, more time is available for planning therapies for life-threatening diseases and for preparing as well as publishing research.

About the author
Elif Karakurt
medical content creator
Elif is a medical student and works for Cytolytics in the branches of content creation and marketing alongside her studies. She is the head of the Cytolytics blog and could already gather experience in writing medical articles for various magazines. Her interests are recent health issues and news about medicine, health technologies, and digital health.