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Mastering the Challenge of Fluorescence Intensity Calibration

  • The calibration of fluorescence signals in assays as well as in biological systems is a key requirement. Especially for quantitative studies of living cells, e.g. expression of biomolecules the generation of concentration-proportional analytical statements obtained by the fluorescence intensity plays a significant role. This also applies for cell assays like immunofluorescence assays. Also, the determination of local concentrations of target proteins within cells or within their environment is a major challenge in modern biology. Proper calibration of the fluorescence imaging systems is the prerequisite to ensure that test results from different instruments and different test environments are comparable. The existing calibration solutions for fluorescence imaging systems, however, mostly depend on simple instrument calibration without reference to absolute physical standards or with insufficient stability for multiple measurements. In order to obtain the necessary quantitativeThe calibration of fluorescence signals in assays as well as in biological systems is a key requirement. Especially for quantitative studies of living cells, e.g. expression of biomolecules the generation of concentration-proportional analytical statements obtained by the fluorescence intensity plays a significant role. This also applies for cell assays like immunofluorescence assays. Also, the determination of local concentrations of target proteins within cells or within their environment is a major challenge in modern biology. Proper calibration of the fluorescence imaging systems is the prerequisite to ensure that test results from different instruments and different test environments are comparable. The existing calibration solutions for fluorescence imaging systems, however, mostly depend on simple instrument calibration without reference to absolute physical standards or with insufficient stability for multiple measurements. In order to obtain the necessary quantitative information, it is required to calibrate both instruments and the corresponding assays, ideally with one calibrating system. We present several new calibration tools for the most commonly used platforms for cytometry, i.e. fluorescence microscope based systems and flow cytometry based systems. They are comprised of ultra-stable fluorophores that are encapsulated in bio- and cell-compatible polymer matrices. Each tool is tailored to the specific requirements of the different read-out platforms. Both re-usable single-color calibration slides for fluorescence microscopes as well as new multi-color calibration beads are presented. The traceability of the calibration tools to international standards and the wide calibration range of the fluorescence intensity are illustrated. They are thus suitable for calibration of the signal over concentration and quantum yield to that of the targets.zeige mehrzeige weniger

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Metadaten
Autoren/innen:T. Thiele, H. Borcherding, Peter Carl, U. Schedler
Dokumenttyp:Posterpräsentation
Veröffentlichungsform:Präsentation
Sprache:Englisch
Jahr der Erstveröffentlichung:2016
Organisationseinheit der BAM:1 Analytische Chemie; Referenzmaterialien
1 Analytische Chemie; Referenzmaterialien / 1.8 Umweltanalytik
DDC-Klassifikation:Naturwissenschaften und Mathematik / Chemie / Analytische Chemie
Freie Schlagwörter:Calibration; Flow Cytometry; Fluoresence
Themenfelder/Aktivitätsfelder der BAM:Analytical Sciences
Analytical Sciences / Qualitätssicherung
Veranstaltung:DGFZ 26th Annual Meeting
Veranstaltungsort:Berlin, Germany
Beginndatum der Veranstaltung:05.10.2016
Enddatum der Veranstaltung:07.10.2016
Verfügbarkeit des Dokuments:Datei im Netzwerk der BAM verfügbar ("Closed Access")
Datum der Freischaltung:11.10.2016
Referierte Publikation:Nein