Application of TOF-SIMS imaging to C. elgans

• Project/Area Number: 25440082
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Research Field: Cell biology
• Research Institution: Kanagawa University of Human Services (2015); Hamamatsu University School of Medicine (2013-2014)
• Principal Investigator: Kimura Yoshishige 神奈川県立保健福祉大学, 保健福祉学部, 准教授 (90274703)
• Co-Investigator(Renkei-kenkyūsha): SETOU Mitsutoshi 浜松医科大学, 医学部, 教授 (20302664)
• Research Collaborator: HAMEED Saira ; GOTO Kensuke ; ISHIZAKI Itsuko ; SANADA Noriaki ; SUZUKI Mayumi
• Project Period (FY): 2013-04-01 – 2016-03-31
• Project Status: Completed (Fiscal Year 2015)
• Budget Amount: ¥5,200,000 (Direct Cost: ¥4,000,000、Indirect Cost: ¥1,200,000); Fiscal Year 2015: ¥1,040,000 (Direct Cost: ¥800,000、Indirect Cost: ¥240,000); Fiscal Year 2014: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000); Fiscal Year 2013: ¥2,600,000 (Direct Cost: ¥2,000,000、Indirect Cost: ¥600,000)
• Keywords: 質量分析イメージング法 / TOF-SIMS / 線虫 / メタボローム / イメージングマススペクトロメトリー / メタボローム解析

Outline of Final Research Achievements

Imaging mass spectrometry (IMS) is a two-dimensional mass spectrometry to visualize the spatial distribution of biomolecules, which does not need either separation or purification of target molecules. In this study, our goal is to reveal the cellar distribution of biomolecules of C. elegans (a commonly used model organism for the life science) by application of IMS approach. Since C. elegans is a small animal, we have used TRIFT V, TOF-SIMS (Time-of-Flight Secondary Mass Spectrometry) system (ULVAC-PHI, Inc), which enables us to obtain nanoscale microstructure. By comparison of several sample preparation methods, the frozen sections of C. elegans fixed by paraformaldehyde (PFA) were most suitable for TOF-SIMS analysis. By sputtering of Ar gas cluster ion beam (Ar-GCIB), the sensitivity to fatty acids (e.g. stearic acid (SA), oleic acid (OA), and eicosapentaenoic acid(EPA)) was significantly enhanced, and high resolution images of biomolecules were successfully acquired.

Research Products

• [Journal Article] Direct profiling of the phospholipid composition of adult Caenorhabditis elegans using whole-body imaging mass spectrometry. (2015)
  • Author(s): Hameed S, Ikegami K, Sugiyama E, Matsushita S, Kimura Y, Hayasaka T, Sugiura Y, Masaki N, Waki M, Ohta I, Hossen MA, Setou M
  • Journal Title: Anal Bioanal Chem Volume: 407 Issue: 25 Pages: 7589-7602
  • DOI: 10.1007/s00216-015-8932-7
  • Peer Reviewed / Open Access / Int'l Joint Research / Acknowledgement Compliant
• [Presentation] Intraflagellar transport in sensory cilia is tuned by environmental stimuli by tubulin polyglutamylation. (2013)
  • Author(s): Kimura, Y. Konno, A., Tsutsumi, K., Hameed, S., Ikegami, K., Setou, M.
  • Organizer: 北米細胞生物学会
  • Place of Presentation: ニューオーリンズ(米国)
• [Presentation] Environmental stimulus-responsive behavioral plasticity is regulated by intraflagellar transport of sensory cilia. (2013)
  • Author(s): Kimura, Y., Tsutsumi, K., Konno, A., Ikegami, K., Hameed, S, Kaplan, O.I., Teramoto,T., Fujiwara, M., Kunitomo, H., Ishihara, K., Iino, Y., Blacque, O.E., Setou, M.
  • Organizer: FASEB Meeting ----Biology of Cilia and Flagella
  • Place of Presentation: ナイアガラフォールズ(米国)
• [Presentation] Starvation-responsive behavioral plasticity is tuned by tubulin polyglutamylation in sensory cilia (2013)
  • Author(s): Kimura, Y., Tsutsumi, K., Konno, A., Ikegami, K., Hameed, S, Kaplan, O.I., Teramoto,T., Fujiwara, M., Kunitomo, H., Ishihara, K., Iino, Y., Blacque, O.E., Setou, M
  • Organizer: 国際線虫ミーティング
  • Place of Presentation: ロサンゼルス(米国)
• [Presentation] Environmental responsiveness in intraflagellar transport is regulated by tubulin polyglutamylation. (2013)
  • Author(s): 木村芳滋, 紺野在, Saira Hameed, 瀬藤光利
  • Organizer: 日本細胞生物学会
  • Place of Presentation: 名古屋