Quantitative characterization of single cells by use of immunocytochemistry combined with multiplex LA-ICP-MS.
Actual research demonstrates that LA-ICP-MS is capable of being used as an imaging tool with cellular resolution. The aim of this investigation was the method development for LA-ICP-MS to extend the versatility to quantitative and multiplexing imaging of single eukaryotic cells. For visualization of individual cells selected, lanthanide-labeled antibodies were optimized for immuno-imaging of single cells with LA-ICP-MS. The molar content of the artificial introduced labels per cell was quantified using self-made nitrocellulose-coated slides for matrix-matched calibration and calculated amounts were in the range of 3.1 to 17.8 atmol per cell. Furthermore, the quantification strategy allows a conversion of 2D intensity profiles based on counts per second (cps) to quantitative 2D profiles representing the molar amount of the artificial introduced elemental probes per pixel for each individual cell. Graphical abstract ᅟ.
Bundesanstalt für Materialforschung und -prüfung (BAM), Richard-Willstätter-Str. 11, 12489, Berlin, Germany.
Larissa
L
Mueller
Bundesanstalt für Materialforschung und -prüfung (BAM), Richard-Willstätter-Str. 11, 12489, Berlin, Germany.|Institut für Chemie, Humboldt-Universität zu Berlin, Brook-Taylor-Straße 2, 12489, Berlin, Germany.
Antje J
AJ
Herrmann
Bundesanstalt für Materialforschung und -prüfung (BAM), Richard-Willstätter-Str. 11, 12489, Berlin, Germany.
Sandra
S
Techritz
Bundesanstalt für Materialforschung und -prüfung (BAM), Richard-Willstätter-Str. 11, 12489, Berlin, Germany.|Institut für Chemie, Humboldt-Universität zu Berlin, Brook-Taylor-Straße 2, 12489, Berlin, Germany.
Ulrich
U
Panne
Bundesanstalt für Materialforschung und -prüfung (BAM), Richard-Willstätter-Str. 11, 12489, Berlin, Germany. Norbert.Jakubowski@bam.de.
Norbert
N
Jakubowski
D016475:3T3 Cells; D000818:Animals; D000906:Antibodies / Q000032:analysis; D005347:Fibroblasts / Q000166:cytology*; D007150:Immunohistochemistry / Q000379:methods*; D028581:Lanthanoid Series Elements / Q000032:analysis; D013058:Mass Spectrometry / Q000379:methods*; D051379:Mice; D059010:Single-Cell Analysis / Q000379:methods*; D013194:Staining and Labeling / Q000379:methods
D000906:Antibodies; D028581:Lanthanoid Series Elements
Cell systems/single cell analysis; ICP; Immunoassays/ELISA; Laser ablation