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, ESI,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.…
Autor*innen: | Konrad Loehr |
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Koautor*innen: | Ulrich PanneORCiD, Norbert JakubowskiORCiD, Robert Wellhausen |
Dokumenttyp: | Vortrag |
Veröffentlichungsform: | Präsentation |
Sprache: | Englisch |
Jahr der Erstveröffentlichung: | 2017 |
DDC-Klassifikation: | Naturwissenschaften und Mathematik / Chemie / Analytische Chemie |
Freie Schlagwörter: | Single cell arraying |
Veranstaltung: | Metallomics 2017 |
Veranstaltungsort: | Vienna, Austria |
Beginndatum der Veranstaltung: | 13.08.2017 |
Enddatum der Veranstaltung: | 17.08.2017 |
Verfügbarkeit des Dokuments: | Datei im Netzwerk der BAM verfügbar ("Closed Access") |
Datum der Freischaltung: | 08.09.2017 |
Referierte Publikation: | Nein |
Eingeladener Vortrag: | Nein |