Guidelines for the use of flow cytometry and cell sorting in immunological studies (third edition)

The third edition of Flow Cytometry Guidelines provides the key aspects to consider when performing flow cytometry experiments and includes comprehensive sections describing phenotypes and functional assays of all major human and murine immune cell subsets. Notably, the Guidelines contain helpful tables highlighting phenotypes and key differences between human and murine cells. Another useful feature of this edition is the flow cytometry analysis of clinical samples with examples of flow cytometry applications in the context of autoimmune diseases, cancers as well as acute and chronic infectious diseases. Furthermore, there are sections detailing tips, tricks and pitfalls to avoid. All sections are written and peer‐reviewed by leading flow cytometry experts and immunologists, making this edition an essential and state‐of‐the‐art handbook for basic and clinical researchers.DFG, 389687267, Kompartimentalisierung, Aufrechterhaltung und Reaktivierung humaner Gedächtnis-T-Lymphozyten aus Knochenmark und peripherem BlutDFG, 80750187, SFB 841: Leberentzündungen: Infektion, Immunregulation und KonsequenzenEC/H2020/800924/EU/International Cancer Research Fellowships - 2/iCARE-2DFG, 252623821, Die Rolle von follikulären T-Helferzellen in T-Helferzell-Differenzierung, Funktion und PlastizitätDFG, 390873048, EXC 2151: ImmunoSensation2 - the immune sensory syste

Molekulare Proteinwechselwirkungen innerhalb eines Photosignalkomplexes in Membranen - untersucht mit fluoreszenzspektroskopischen Techniken

During the past few years detailed knowledge of the structure of sensory rhodopsin II (NpSRII) and its cognate transducer HtrII from Natronobacterium pharaonis has been gained. NpSRII, a member of the rhodopsin family, serves as a model protein for G-protein coupled receptors. To increase the state of knowledge of the relations between structure and function, earlier studies determined the strength of NpSRII/NpHtrII-binding in detergent buffer. However, little is known about the complex formation in more physiological conditions like in lipid membranes. Binding studies for low protein concentrations as they occur in N. pharaonis have not been performed to this date. The evaluation of NpSRII/NpHtrII-binding was implemented by measuring Förster resonance energy transfer (FRET) of fluorescently labelled proteins at different concentrations. In this work the dissociation constant of NpSRII/NpHtrII in detergent buffer (KD = 200 nM) was reproduced and the complex binding in large unilamellar vesicles (LUV) was analyzed, subsequently. A comparison of FRET-efficiencies, as observed for proteins in detergent buffer and in lipid membranes, shows increased binding affinities of NpSRII and NpHtrII in lipids of at least one order of magnitude. Hence, these results support previous studies, which show that the results observed in detergent buffers are limited in their informational value. In further studies the proteoliposomes were incorporated into giant unilamellar vesicles (GUV) by peptide-induced membrane fusion, to mimic physiologically low protein concentrations. This led to a molar protein-lipid-ratio of 1:1,000,000, as it occurs in bacteria. From this model system exact diffusion coefficients of fluorescently labelled NpSRII and NpHtrII were determined by using the new 2fFCS-technique (two-focus fluorescence correlation spectroscopy). The results indicate, that the NpSRII/NpHtrIIbinding is observed even at physiologically protein concentration. The diffusion coefficients were proportional to the inverse cylinder radius of the proteins. This is in the line with recently published studies that show discrepancies to the far spread model of Saffman and Delbrück

Guidelines for the use of flow cytometry and cell sorting in immunological studies (third edition)

The third edition of Flow Cytometry Guidelines provides the key aspects to consider when performing flow cytometry experiments and includes comprehensive sections describing phenotypes and functional assays of all major human and murine immune cell subsets. Notably, the Guidelines contain helpful tables highlighting phenotypes and key differences between human and murine cells. Another useful feature of this edition is the flow cytometry analysis of clinical samples with examples of flow cytometry applications in the context of autoimmune diseases, cancers as well as acute and chronic infectious diseases. Furthermore, there are sections detailing tips, tricks and pitfalls to avoid. All sections are written and peer-reviewed by leading flow cytometry experts and immunologists, making this edition an essential and state-of-the-art handbook for basic and clinical researchers.ISSN:0014-2980ISSN:1521-414

Guidelines for the use of flow cytometry and cell sorting in immunological studies (third edition)

Cossarizza A, Chang H‐D, Radbruch A, et al. Guidelines for the use of flow cytometry and cell sorting in immunological studies (third edition). European Journal of Immunology. 2021;51(12):2708-3145.The third edition of Flow Cytometry Guidelines provides the key aspects to consider when performing flow cytometry experiments and includes comprehensive sections describing phenotypes and functional assays of all major human and murine immune cell subsets. Notably, the Guidelines contain helpful tables highlighting phenotypes and key differences between human and murine cells. Another useful feature of this edition is the flow cytometry analysis of clinical samples with examples of flow cytometry applications in the context of autoimmune diseases, cancers as well as acute and chronic infectious diseases. Furthermore, there are sections detailing tips, tricks and pitfalls to avoid. All sections are written and peer-reviewed by leading flow cytometry experts and immunologists, making this edition an essential and state-of-the-art handbook for basic and clinical researchers

Guidelines for the use of flow cytometry and cell sorting in immunological studies (third edition)

The third edition of Flow Cytometry Guidelines provides the key aspects to consider when performing flow cytometry experiments and includes comprehensive sections describing phenotypes and functional assays of all major human and murine immune cell subsets. Notably, the Guidelines contain helpful tables highlighting phenotypes and key differences between human and murine cells. Another useful feature of this edition is the flow cytometry analysis of clinical samples and respective applications of flow cytometry in the context of a variety of autoimmune diseases, cancers as well as acute and chronic infectious diseases such as COVID-19. Furthermore, there are sections detailing tips, tricks and pitfalls to avoid. All sections are written and peer-reviewed by leading flow cytometry experts and immunologists, making this edition an essential and state-of-the-art handbook for basic and clinical researchers