Extended Data Fig. 2: Screen for differentiation markers of cell populations for FACS-based sorting.
From: Complex mammalian-like haematopoietic system found in a colonial chordate
a, b, Examples of antibodies screened by CyTOF mass cytometry with B. schlosseri cells analysed by 2D mass spectrometry. The CyTOF screen was performed once. a, Examples of antibodies considered as nonspecific binders because different antibodies showed the same binding patterns. b, Examples of antibodies considered as specific binders because a cell population was bound by one antibody but not the other (red rectangle). c, d, Examples of validation screen by flow cytometry of antibodies with specific binding by CyTOF, done in 2D fluorescence excited by the same laser owing to autofluorescence. Negative FACS validation was done twice and positive three times. c, Negative validation of CCR2 labelled with PE. d, Positive validation of CD57 labelled with Pacific Blue. e, Flow cytometry of live B. schlosseri cells labelled with anti-BHF mouse serum. Anti-mouse Alexa Fluor-647 was used as a secondary antibody. f, Example of positive differential labelling by the lectin concanavalin-A in PE-Cy7 by FACS. The experiment was performed three times. g, Confocal imagery of membrane concanavalin-A labelling with Alexa Fluor-633 in red and Hoechst DNA labelling in blue. The experiment was performed once. Scale bar, 20 µm.
