Abstract
Recent advances in the field of flow cytometry (FCM) have highlighted the importance of incorporating it as a basic analysis tool in laboratories. FCM not only allows the identification of cell subpopulations by detecting the expression of molecules in the cell membrane or cytoplasm, but it can also quantify and identify soluble molecules. The proper functioning of the FCM requires six fundamental systems, from those related to the transport of events to the systems dedicated to the analysis of information. In this review, we have identified the main considerations that every FCM user must know for an optimal antibody panel design, the quality systems that must govern the FCM protocols to guarantee reproducible results in research or clinical laboratories. Finally, we have introduced the current evidence that highlights the relevance of FCM in the investigation and clinical diagnosis of respiratory diseases, establishing important advances in the basic and clinical study of diseases as old as Tuberculosis along with the recent proposals for the monitoring and classification of patients infected with the new SARS-CoV2 virus.
Highlights
The quantitative analysis of blood cells began in 1873 with the development of the hemocytometer, designed by Louis–Charles Malassez; subsequently, the cellular photoelectric counters made it possible to identify “individual events” smaller than 10 microns in single-cell suspension [1,2,3]
For the development of Flow cytometry (FCM), the inkjet system of the printers was incorporated into the Coulter technology, allowing to generate a system of micro-drops in continuous fluid, which favoured the basic principle of cell separation known as Fluorescence-Activated Cell Sorting (FACS) [7,8]
Fluorescence minus one (FMO): These controls contain all the monoclonal antibodies (mAb)-F used in the panel, except for one, which is relevant to the molecular markers to be studied
Summary
The quantitative analysis of blood cells began in 1873 with the development of the hemocytometer, designed by Louis–Charles Malassez; subsequently, the cellular photoelectric counters made it possible to identify “individual events” smaller than 10 microns in single-cell suspension [1,2,3]. Crossland-Taylor developed the “hydrodynamic focusing” technology, which allowed automated systems for greater speed of analysis in cell counting; and the detection system based on the electrical conductance of cells suspended in a conductive fluid, developed by Wallace H. For the development of FCM, the inkjet system of the printers was incorporated into the Coulter technology, allowing to generate a system of micro-drops in continuous fluid, which favoured the basic principle of cell separation known as Fluorescence-Activated Cell Sorting (FACS) [7,8]. Clinical CytomCeutrrryenStlyo,citehtey “(IInCteCrnSa)t”io,naalreStohcieetyinsfotirtuAtdiovnanscetmhaenttdoicftaCtyetotmheetrcyor(rISeActC)s”taannddarthdeization of techniques“aInntderenqatuioipnaml eCnlitnficoarl FCCyMtom, teotrgyeSnoecrieattye (eIffCeCcSt)i”v, earaendthreeipnrsotidtuuticoinbslethraetsudlictsta.teFCthMe choarsreectstablished itself as onsFetCaonMdf ahtrhadseizeasmttaioabnilinsohftetoedcoihtlnsseiqltfuoaesssouannpedpoeofqrtuhtipetmmheeanindt fitooarogFlnsCtoMos,situsopagpenondretrtachtleeinedfiifcaegacntliovmseisaonandnidtroecprlirinonidcgaulcomibfloednriietsoseurailntssge. s since it allows the omf doinseitaosersinsigncoefittahlelowexspthreesmsoionintoorifngspoef cthifie ecxcperellssmionarokfesprsec(iFficigcuelrlem1aArke).rs (Figure 1A)
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