Quantitative LA-ICP-MS of single cells: Comparison of single spot ablation and imaging

  • LA-ICP-MS finds widespread use in the analysis of biological samples and its importance as an analysitical method is still increasing. Often tissues or single cells are ablated in imaging mode on glass slides, but the analysis time increases by reducing the spot size of the laser. Additionally, with increased lateral resolution a loss in sensitivity is inherently connected and this holds true in particular for single cell analysis in imaging mode. However, if only the composition of elements in a single cell needs to be measured, complete ablation of cells by single shots looks promising. This approach was already discussed by Managh et al. and will be discussed in more detail in this presentation. The aim of this study is to develop a calibration strategy for single cell analysis based on the total consumption concept. It will be shown that this concept provides quantitative results at single cell level, if the target material is ablated. We compare two different laser spot sizes, 30LA-ICP-MS finds widespread use in the analysis of biological samples and its importance as an analysitical method is still increasing. Often tissues or single cells are ablated in imaging mode on glass slides, but the analysis time increases by reducing the spot size of the laser. Additionally, with increased lateral resolution a loss in sensitivity is inherently connected and this holds true in particular for single cell analysis in imaging mode. However, if only the composition of elements in a single cell needs to be measured, complete ablation of cells by single shots looks promising. This approach was already discussed by Managh et al. and will be discussed in more detail in this presentation. The aim of this study is to develop a calibration strategy for single cell analysis based on the total consumption concept. It will be shown that this concept provides quantitative results at single cell level, if the target material is ablated. We compare two different laser spot sizes, 30 µm and 110 µm. The smaller spot size of 30 µm is used for imaging and the larger spot size of 110 µm is used for the targeted ablation of complete single cells. Cells were stained by two novel cell dyes, mDOTA-Tb (CheMatech, Dijon, France), which binds to free thiol groups of proteins, and Ir-DNA intercalator (Maxpar®, Fluidigm, San Francisco, USA), which stains DNA quantitatively. Finally, quantification of Ir and Tb will be performed by matrix matching calibration, following the approach of Drescher et al.[1], i.e. spotting of a dilution series of Ir and Tb standards on nitrocellulose membrane. Results obtained so far show a good correlation between imaging and single spot ablation, indicating that the total consumption concept may serve as an acceptable assumption.zeige mehrzeige weniger

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Metadaten
Autor*innen:Konrad Loehr
Koautor*innen:Antje Jutta Herrmann, Heike Traub, Norbert Jakubowski
Dokumenttyp:Vortrag
Veröffentlichungsform:Präsentation
Sprache:Englisch
Jahr der Erstveröffentlichung:2017
DDC-Klassifikation:Naturwissenschaften und Mathematik / Chemie / Analytische Chemie
Freie Schlagwörter:Bioimaging; LA-ICP-MS; Single cell analysis; Single spot ablation
Veranstaltung:EWCPS
Veranstaltungsort:St. Anton am Arlberg, Austria
Beginndatum der Veranstaltung:19.02.2017
Enddatum der Veranstaltung:24.02.2017
Verfügbarkeit des Dokuments:Datei im Netzwerk der BAM verfügbar ("Closed Access")
Datum der Freischaltung:12.04.2017
Referierte Publikation:Nein
Eingeladener Vortrag:Nein
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