rabbit polyclonal igg anti-pax6  (Covance)

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

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: Negative 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 negative ion mode of 60-day-old brain organoids detected 168 lipids across the measured mass range of m/z 200-1,000. This includes fatty acids (m/z 200–600) and phospholipids (m/z 700–1,000) lipid signatures in negative 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). Arrows point to rosettes. (C) Ion distribution images of fatty acids and other lipids that are visualized and detected in negative ion mode represent several lipid classes including fatty acids and glycerophospholipids. All images were generated with a bin width of Δm/z = 0.01 at a mass resolving power of 70,000 at m/z = 200 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%. CerPE, ceramide-phosphoethanolamine; PA, phosphatidic acid; PE, phosphatidyl-ethanolamine; PS, phosphatidyl-serine; PI, phosphatidyl-inositol.

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

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: Sphingolipid CerPE 36:1; O2 is selectively abundant in rosettes, while PE 38:3 is selectively absent from rosettes. Positive (A) and negative (B) ion mode shows overlaid distribution of CerPE 36:1; O2 and PE 38:3 in two 60-day-old brain organoids. Confocal micrographs of sections adjacent to the sections used for MSI show expression of DAPI (nuclear stain), PAX6 (marker of neuroprogenitors localized in rosettes) and doublecortin (DCX, marker of immature neurons). Inset in (B) is a high magnification of a rosette, showing expression of PA×6 and not DCX.

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: Expressing, Staining, Marker