Live single-cell video-mass spectrometry for cellular and subcellular molecular detection and cell classification.
The molecular content from the cytoplasm of a live, single mammalian cell and its organelle were trapped with a nano-electrospray ionization (ESI) tip acting as a micropipette under a video microscope, and hundreds of small molecular peaks were detected by direct nano-ESI mass spectrometry (MS). Granule- or cytoplasm-specific peaks in a mast cell (RBL 2H3) model were extracted by paired t-test to demonstrate their specific localization. Some of the typical and specific molecules were successfully identified by MS/MS analysis. This method was also applied to the cell classification of seven types of cell lines at the single-cellular level by principal component analysis (PCA), revealing seven clusters in the multivariate score plot.
Mizuno
Hajime
H
Graduate School of Biomedical Sciences, Hiroshima University, 1-2-3 Kasumi, Minami-ku, Hiroshima City 734-8551, Japan.
Tsuyama
Naohiro
N
Harada
Takanori
T
Masujima
Tsutomu
T
D000818:Animals; D002478:Cells, Cultured; D003593:Cytoplasm / Q000737:chemistry* / Q000648:ultrastructure; D006801:Humans; D051379:Mice; D008853:Microscopy / Q000379:methods; D015388:Organelles / Q000737:chemistry* / Q000648:ultrastructure; D051381:Rats; D021241:Spectrometry, Mass, Electrospray Ionization / Q000379:methods; D053719:Tandem Mass Spectrometry / Q000379:methods