Towards quantitative high throughput single cell LA-ICP-MS: microarraying of single cells and calibration standards via piezo based non-contact dispensing

  • Analysis of single cells via LA-ICP-MS is a technique with great potential, however manual targeting of single cells is laborious and therefore microarraying of cells looks promising. In this work, we investigate the potential of a commercial non-contact piezo dispenser arraying system (S3, Scienion AG, Berlin), equipped with a novel technology for single-cell Isolation called CellenONE™ (Cellenion, Lyon). Usually if one aims to create a microarray of single cells via spotting a suitably diluted cell suspension, one will observe a Poisson-distributed cell number per spot. CellenONE™ overcomes this problem by controlling the number of cells optically in the piezo dispense capillary (PDC) via image recognition to obtain true single cell arrays. We will present the figures of merit of the customized and optimized setup. As a model system we use THP-1 cells stained with two dyes, mDOTAHo(CheMatech, Dijon), and Ir-DNA intercalator (Fluidigm, San Francisco) and used LA-ICP-MS (NWR213,Analysis of single cells via LA-ICP-MS is a technique with great potential, however manual targeting of single cells is laborious and therefore microarraying of cells looks promising. In this work, we investigate the potential of a commercial non-contact piezo dispenser arraying system (S3, Scienion AG, Berlin), equipped with a novel technology for single-cell Isolation called CellenONE™ (Cellenion, Lyon). Usually if one aims to create a microarray of single cells via spotting a suitably diluted cell suspension, one will observe a Poisson-distributed cell number per spot. CellenONE™ overcomes this problem by controlling the number of cells optically in the piezo dispense capillary (PDC) via image recognition to obtain true single cell arrays. We will present the figures of merit of the customized and optimized setup. As a model system we use THP-1 cells stained with two dyes, mDOTAHo(CheMatech, Dijon), and Ir-DNA intercalator (Fluidigm, San Francisco) and used LA-ICP-MS (NWR213, ESI, Portland; ElementXR, ThermoScientific, Bremen) to measure Ho and Ir signals from single cells. Both metallo-dyes will be quantified by matrix matched calibration at single cell resolution after Wang et al. (Anal. Chem. 2014, 86, 10252−10256). We believe that this novel approach opens new ways for automated quantitative single cell LA-ICP-MS.zeige mehrzeige weniger

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Metadaten
Autoren/innen:Konrad Loehr
Koautoren/innen:Ulrich Panne, Norbert Jakubowski, Robert Wellhausen
Dokumenttyp:Vortrag
Veröffentlichungsform:Präsentation
Sprache:Englisch
Jahr der Erstveröffentlichung:2017
Organisationseinheit der BAM:1 Analytische Chemie; Referenzmaterialien
1 Analytische Chemie; Referenzmaterialien / 1.1 Anorganische Spurenanalytik
DDC-Klassifikation:Naturwissenschaften und Mathematik / Chemie / Analytische Chemie
Freie Schlagwörter:Single cell arraying
Themenfelder/Aktivitätsfelder der BAM:Analytical Sciences
Veranstaltung:Metallomics 2017
Veranstaltungsort:Vienna, Austria
Beginndatum der Veranstaltung:13.08.2017
Enddatum der Veranstaltung:17.08.2017
Verfügbarkeit des Volltexts:Volltext-PDF im Netzwerk der BAM verfügbar ("Closed Access")
Datum der Freischaltung:08.09.2017
Referierte Publikation:Nein