Lifetime Barcoding of Polystyrene Beads with Fluorescent Nanocrystals for Fluorescent Lifetime Detection in Flow Cytometry
- Multiplexed encoding schemes of nano- and micrometer sized polymer particles with fluorescent dyes or quantum dots (QDs) and their optical detection, are of increasing interest for applications in the life sciences, for example in flow cytometry. Almost all strategies utilizing fluorescence focus on spectrally distinguishable emission bands or colors and different intensity levels as fluorescence codes. In this work the goal is to perform multiplexing with encoding fluorophores with different fluorescence lifetimes (LTs). In comparison to the spectral multiplexing strategies this has the advantage, that the different fluorescence LT codes can be measured with the same excitation and emission wavelength, thus reducing instrumental costs. Moreover, LTs should not depend on emitter concentration. Unlike organic dyes, the LTs of which are typically <10ns, the fluorescence LTs of ternary semiconductor QDs which represent a “green” alternative to conventional Cd-containing QDs are in theMultiplexed encoding schemes of nano- and micrometer sized polymer particles with fluorescent dyes or quantum dots (QDs) and their optical detection, are of increasing interest for applications in the life sciences, for example in flow cytometry. Almost all strategies utilizing fluorescence focus on spectrally distinguishable emission bands or colors and different intensity levels as fluorescence codes. In this work the goal is to perform multiplexing with encoding fluorophores with different fluorescence lifetimes (LTs). In comparison to the spectral multiplexing strategies this has the advantage, that the different fluorescence LT codes can be measured with the same excitation and emission wavelength, thus reducing instrumental costs. Moreover, LTs should not depend on emitter concentration. Unlike organic dyes, the LTs of which are typically <10ns, the fluorescence LTs of ternary semiconductor QDs which represent a “green” alternative to conventional Cd-containing QDs are in the range of several hundred ns, independent of oxygen concentration, and can be tuned to a certain extent by chemical composition and surface chemistry. This presents a time region that can barely be covered by other emitters that have either much shorter or longer lifetimes. In this project, different encoding strategies will be assessed, and the encoded particles will then be used for fluorescence assays for the analysis of several targets in parallel. Therefore, the encoded particles will be functionalized with different target-specific bioligands and read out with a specifically designed flow cytometer enabling time-resolved fluorescence detection. With this instrument, the particles will be discriminated by their fluorescence LTs in one detection channel while the analytes will be quantified by fluorescence labels in a second channel in the intensity domain.…
Autor*innen: | Oskar Hübner |
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Koautor*innen: | Ute Resch-GengerORCiD, Karl David WegnerORCiD, Kristin Hoffmann |
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: | AIS; Assay; Barcoding; Bead; Dye; Encoding; Flow cytometry; Fluorescence; InP; Lifetime; Method; Multiplexing; Quantum dot |
Themenfelder/Aktivitätsfelder der BAM: | Chemie und Prozesstechnik |
Material | |
Material / Nano | |
Veranstaltung: | Tag der Chemie 2019 |
Veranstaltungsort: | Berlin, Germany |
Beginndatum der Veranstaltung: | 11.07.2019 |
Enddatum der Veranstaltung: | 11.07.2019 |
Verfügbarkeit des Dokuments: | Datei im Netzwerk der BAM verfügbar ("Closed Access") |
Datum der Freischaltung: | 27.11.2019 |
Referierte Publikation: | Nein |