Fig. 4: Mass spectrometry imaging of single cell cultures at sub-cellular spatial resolutions.

a Neuroblastoma cell line (SH-SY5Y) imaged through t-MALDI-P MSI at different pixel sizes. MSI pixel sizes of (left-to-right) 5 µm, 2 µm, 1.5 µm and 1 µm display the overlaid ion-distribution images annotated within the first panel. b Osteosarcoma cell line (U2OS) stained with DAPI nuclear stain and the lipid stain, Nile red, and imaged through MSI at 1 µm (top) and 3-channel (390/470 nm, 555/550 nm, 475/510 nm) 40× fluorescence microscopy (bottom). c Neuroblastoma cell line (SH-SY5Y) stained with the nuclear stain Hoechst and mitochondrial stain MitoTracker Deep Red and imaged through MSI at 1.5 µm (top) and 2-channel (390/470 nm, 635/700 nm) 40× fluorescence microscopy (bottom). d Established patient derived cancer cell lines, where monocultures of each line were stained with different fluorescent CellBrite membrane dyes (red, green, blue) before co-culture. (Top) MSI at 750 nm pixel size, and (bottom) 3-channel (390/470 nm, 555/550 nm, 475/510 nm) 10× fluorescence microscopy. Mass spectral investigations for each single cell MSI can be found in Supplementary Fig. 14. All microscopy images were obtained in triplicate (n = 3) and display similar results.