Journal: Frontiers in Molecular Biosciences
Article Title: Mass spectrometry imaging as an emerging tool for studying metabolism in human brain organoids
doi: 10.3389/fmolb.2023.1181965
Figure Lengend Snippet: Positive ion mode MSI from 60-day-old brain organoids shows different spatial distribution and abundance of lipid species. (A) Average m/z spectra generated from the positive ion mode of 60-day-old brain organoids detected 114 lipid species across the measured mass range of m/z 400-1,000. This included lysolipid (m/z 400–600) and phospholipid (m/z 700–1,000) signals at positive polarity. (B) Hematoxylin and eosin (HE) staining along with confocal micrograph of serial sections shows organoid morphology and expression of DAPI (nuclear stain), PAX6 (marker of neuroprogenitors localized in rosettes) and doublecortin (DCX, marker of immature neurons). Arrow points to a rosette. (C) Ion distribution images of lipids that are visualized and detected in positive ion mode represent several lipid classes including lysolipids, glycerophospholipids and sphingolipids. All images were generated with a bin width of Δm/z = 0.01 at a mass resolving power of 70,000 at m/z = 400 and a mass accuracy within ±5 ppm. The spatial resolution was set to 25 µm/pixel. Each dot on the image is one pixel. The scale bar represents 1 mm and ion intensities are normalized to the total ion count (TIC) and scaled from 0% to 100%. LPC, lysophosphatidyl-choline; PE, phosphatidyl-ethanolamine; PC, phosphocholine; PS, phosphatidyl-serine; SM, sphingomyelin; SHexCer, sulfatide hexosyl ceramide.
Article Snippet: Tissue sections were fixed in 150 μL 4% paraformaldehyde per section for 10 min, before incubation with the primary antibodies diluted in 0.3% Triton-X at 4°C overnight (goat polyclonal IgG DCX sc-271390 (Santa Cruz), rabbit polyclonal IgG PAX6 901301 (Biolegend)).
Techniques: Generated, Staining, Expressing, Marker