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Selective improvement of peptides imaging on tissue by supercritical fluid wash of lipids for matrix-assisted laser desorption/ionization mass spectrometry

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Abstract

There is a high analytical demand for improving the detection sensitivity for various peptides in matrix-assisted laser desorption/ionization imaging mass spectrometry (MALDI-IMS) because exhaustive distribution analyses of various peptides could help to reveal the function of peptides in vivo. To improve the sensitivity of peptide detection, we used supercritical fluid of CO2 (scCO2) as washing solvent for a pretreatment to remove lipids. We evaluated whether our wash method using scCO2 with an entrainer improved the detection of peptides and suppressed lipid detection in MALDI-IMS. Our analysis revealed that the signal intensities of peptides such as m/z 3339.8, 3530.9, 4233.3, 4936.7, and 4963.7 were increased in scCO2-washed samples. The greatest improvement in the signal-to-noise ratio (S/N) was found at m/z 4963.7, which was identified as thymosin β4, with the S/N reaching almost 190-fold higher than the control. Additionally, all of the improved signals were associated with the morphologic structure. Our method allows us to analyze the distribution of molecules, especially in the region of m/z 3000–5200. For these improvements, the polarity difference between scCO2 and the matrix solution used was considered as a key. A wider variety of molecules can be analyzed in the future due to this improvement of the detection sensitivity by optimizing the polarity of scCO2 with various entrainers.

Mass spectra of m/z 4900-5000 obtained from a scCO2-washed tissue (upper, blue) and a control tissue (lower, red). Ion distribution of the signals at m/z 4936.7 and m/z 4963.7 specifically ditected from scCO2-washed samples

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Acknowledgements

The authors thank S. Yamamoto and Y. Nakaya for their technical support with the scCO2 instrument. This work was supported by grants from Japan’s Ministry of Education, Culture, Sports, Science, and Technology (grant nos. 15H05900 and 15H05898B1), a Research Fellowship from the Japan Society for the Promotion of Science (JSPS KAKENHI grant no. JP16J05476), and a grant from Yamada Ryozo Kikin of the Hamamatsu Foundation for Science and Technology Promotion to M.S.

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Authors and Affiliations

  1. Department of Cellular and Molecular Anatomy, Hamamatsu University School of Medicine, 1-20-1 Handayama, Higashi-ku, Hamamatsu, Shizuoka, 431-3192, Japan

    Shoko Matsushita, Noritaka Masaki, Eiji Sugiyama & Mitsutoshi Setou

  2. International Mass Imaging Center, Hamamatsu University School of Medicine, 1-20-1 Handayama, Higashi-ku, Hamamatsu, Shizuoka, 431-3192, Japan

    Shoko Matsushita, Noritaka Masaki, Eiji Sugiyama & Mitsutoshi Setou

  3. Research Fellow of Japan Society for the Promotion of Science, Kojimachi Business Center Building, 5-3-1 Kojimachi, Chiyoda-ku, Tokyo, 102-0083, Japan

    Shoko Matsushita

  4. Department of Applied Chemistry and Biochemical Engineering, Faculty of Engineering, Shizuoka University, 3-5-1 Johoku, Naka-ku, Hamamatsu, Shizuoka, 432-8561, Japan

    Kohei Sato & Nobuyuki Mase

  5. Health Innovation and Technology Center, Faculty of Health Sciences, Hokkaido University, Kita 12, Nishi 5, Kita-ku, Sapporo, Hokkaido, 060-0812, Japan

    Takahiro Hayasaka, Shu-Ping Hui & Hitoshi Chiba

  6. Preeminent Medical Photonics Education and Research Center, Hamamatsu University School of Medicine, 1-20-1 Handayama, Higashi-ku, Hamamatsu, Shizuoka, 431-3192, Japan

    Mitsutoshi Setou

  7. Department of Anatomy, The University of Hong Kong, 6/F, William MW Mong Block 21 Sassoon Road, Pokfulam, Hong Kong, SAR, China

    Mitsutoshi Setou

  8. Riken Center for Molecular Imaging Science, 6-7-3 Minatojima-minamimachi, Chuo-ku, Kobe, Hyogo, 650-0047, Japan

    Mitsutoshi Setou

  9. Division of Neural Systematics, National Institute for Physiological Sciences, 38 Nishigonaka Myodaiji, Okazaki, Aichi, 444-8585, Japan

    Mitsutoshi Setou

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  1. Shoko Matsushita
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  2. Noritaka Masaki
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  3. Kohei Sato
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Correspondence to Mitsutoshi Setou.

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Matsushita, S., Masaki, N., Sato, K. et al. Selective improvement of peptides imaging on tissue by supercritical fluid wash of lipids for matrix-assisted laser desorption/ionization mass spectrometry. Anal Bioanal Chem 409, 1475–1480 (2017). https://doi.org/10.1007/s00216-016-0119-3

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  • DOI: https://doi.org/10.1007/s00216-016-0119-3

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