Preparation and characterization of reference particles for the calibration of flow cytometry for the detection of extracellular vesicles within the EMPIR project MetVes II

  • Extracellular vesicles (EV) are cell-derived particles in body fluids, which have excellent potential as next-generation biomarkers. The exploitation of EV requires reliable measurements, which is currently very difficult, as most EV are smaller than 200 nm. At present, flow cytometry (FCM) is the most appropriate technique for EV analysis in biological samples, as FCM is readily available in many clinical laboratories and allows to identify cell-specific EV at high throughput. However, due to technical variations between different FCM instruments, EV concentration measurements are currently not well comparable between most laboratories. Therefore, EV reference materials and standardized reference methods are urgently needed to calibrate flow rate, light scattering intensity, and fluorescence intensity of FCM in the sub-micrometer size range. This requires a better matching of the optical properties of calibration beads and EV as can be realized with current polystyrene calibrationExtracellular vesicles (EV) are cell-derived particles in body fluids, which have excellent potential as next-generation biomarkers. The exploitation of EV requires reliable measurements, which is currently very difficult, as most EV are smaller than 200 nm. At present, flow cytometry (FCM) is the most appropriate technique for EV analysis in biological samples, as FCM is readily available in many clinical laboratories and allows to identify cell-specific EV at high throughput. However, due to technical variations between different FCM instruments, EV concentration measurements are currently not well comparable between most laboratories. Therefore, EV reference materials and standardized reference methods are urgently needed to calibrate flow rate, light scattering intensity, and fluorescence intensity of FCM in the sub-micrometer size range. This requires a better matching of the optical properties of calibration beads and EV as can be realized with current polystyrene calibration beads. The EMPIR project 18HLT01 “MetVes II” aims to develop synthetic reference materials and traceable measurement methods to standardize EV measurements. The reference materials should resemble EV properties, so that calibrations are reliable and do not require a change of acquisition settings. Hence, the reference materials should contain particles with a traceable number concentration in the range of 109–1012 particles/mL to calibrate flow rate, a traceable size with discrete diameters between 50–1000 nm and a refractive index (RI) in the range of 1.37–1.42 to calibrate scattering intensity, and a traceable fluorescence intensity between 100–100,000 molecules of equivalent soluble fluorochromes (MESF). At BAM, various approaches to prepare such low-RI nanometer-sized reference materials will be studied, preliminary results of the primary characterization of these candidate reference particles will be presented, and possible applications besides FCM-based EV detection will be outlined.zeige mehrzeige weniger

Volltext Dateien herunterladen

  • Poster_2019_Geissler_DGfZ-Tagung.pdf
    eng

Metadaten exportieren

Weitere Dienste

Teilen auf Twitter Suche bei Google Scholar
Metadaten
Autoren/innen:Daniel Geißler
Koautoren/innen:Nithiya Nirmalananthan-Budau, 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
Technik, Medizin, angewandte Wissenschaften / Ingenieurwissenschaften / Ingenieurwissenschaften und zugeordnete Tätigkeiten
Freie Schlagwörter:18HLT01 MetVes II; EMPIR project; Extracellular vesicles (EV); Flow cytometry; Reference particles
Themenfelder/Aktivitätsfelder der BAM:Material
Analytical Sciences
Analytical Sciences / Qualitätssicherung
Material / Nanoskalige Materialien und deren Eigenschaften
Veranstaltung:29th Meeting of the German Society for Cytometry (DGfZ)
Veranstaltungsort:Berlin, Germany
Beginndatum der Veranstaltung:25.09.2019
Enddatum der Veranstaltung:27.09.2019
Verfügbarkeit des Dokuments:Datei im Netzwerk der BAM verfügbar ("Closed Access")
Datum der Freischaltung:13.02.2020
Referierte Publikation:Nein