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Time-resolved flow cytometry

  • The fast identification of a large number of analytes or events is increasingly required in bioanalytical, diagnostic, and security applications. The versatility and straightforward use make multiparametric fluorescence techniques particularly interesting as detection techniques. An established method for high-throughput single-cell and single-particle measurements is flow cytometry (FCM). Using only spectral encoding without further intensity information, state-of-the-art instruments equipped with several light sources and detectors can resolve almost 20 different color codes. However, this is not sufficient to answer complex research questions, e.g. in cell biology and immunology. In contrast, routine applications demand low-cost and sometimes even portable instruments and thus a minimum number of instrument components. Thus, there are currently two main research directions in FCM: the development of methods that can either address increasingly complex analytical challenges orThe fast identification of a large number of analytes or events is increasingly required in bioanalytical, diagnostic, and security applications. The versatility and straightforward use make multiparametric fluorescence techniques particularly interesting as detection techniques. An established method for high-throughput single-cell and single-particle measurements is flow cytometry (FCM). Using only spectral encoding without further intensity information, state-of-the-art instruments equipped with several light sources and detectors can resolve almost 20 different color codes. However, this is not sufficient to answer complex research questions, e.g. in cell biology and immunology. In contrast, routine applications demand low-cost and sometimes even portable instruments and thus a minimum number of instrument components. Thus, there are currently two main research directions in FCM: the development of methods that can either address increasingly complex analytical challenges or provide low-cost and robust approaches for routine multiplex analyses. Common spectral multiplexing approaches face limitations in both directions. On the one hand, spectral overlap of labels restricts the number of codes and makes elaborate correction schemes necessary. On the other hand, even for lower degrees of multiplexing often a sophisticated optical setup is needed. An alternative to spectral multiplexing and intensity encoding is to exploit the luminescence lifetime (LT) as an encoding parameter. This can allow for extending the parameter space in combination with spectral encoding or result in more simple and compact devices due to fewer optical components. The availability of fast electronics enables miniaturized and portable lifetime measurement setups at relatively low cost. LT-FCM requires to master LT determination with a limited number of detected photons due to the short interaction time of the encoded objects with the laser spot. In this study, we address this issue for time-domain cytometry and present a novel lifetime flow cytometry (LT-FCM) platform based on a compact setup and straightforward time-domain measurements utilizing LT-encoded luminescent beads. Moreover, we present the realization of a first bioanalytical assay with LT-encoded beads.zeige mehrzeige weniger

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  • Poster-ANAKON_DK_0204_2019_KH.pdf
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Metadaten
Autoren/innen:Katrin Hoffmann
Koautoren/innen:Daniel Kage, J. Ameskamp, M. Wittkamp, T. Thiele, H. Borcherding, W. Göhde, U. Schedler, Ute Resch-Genger
Dokumenttyp:Posterpräsentation
Veröffentlichungsform:Präsentation
Sprache:Englisch
Jahr der Erstveröffentlichung:2019
Organisationseinheit der BAM:1 Analytische Chemie; Referenzmaterialien
1 Analytische Chemie; Referenzmaterialien / 1.2 Biophotonik
DDC-Klassifikation:Naturwissenschaften und Mathematik / Chemie / Analytische Chemie
Freie Schlagwörter:Bead-based assays; Flow cytometry; Fluorescence; Lifetime-encoding; Time-resolved
Themenfelder/Aktivitätsfelder der BAM:Analytical Sciences
Analytical Sciences / Zerstörungsfreie Prüfung und Spektroskopie
Veranstaltung:ANAKON 2019
Veranstaltungsort:Münster, Germany
Beginndatum der Veranstaltung:25.03.2019
Enddatum der Veranstaltung:28.03.2019
Verfügbarkeit des Volltexts:Volltext-PDF im Netzwerk der BAM verfügbar ("Closed Access")
Datum der Freischaltung:03.04.2019
Referierte Publikation:Nein