Development of sample preparation for single cell 3D mass spectrometry imaging

2022 Fiscal Year Final Research Report

• Project/Area Number: 19K12754
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Review Section: Basic Section 90110:Biomedical engineering-related
• Research Institution: National Institute of Genetics
• Principal Investigator: Masaki Noritaka 国立遺伝学研究所, 遺伝メカニズム研究系, 特任研究員 (80614073)
• Project Period (FY): 2019-04-01 – 2023-03-31
• Keywords: 質量分析 / 質量分析イメージング / TOF-SIMS / 脂質分析 / 脂肪酸 / 脂肪滴 / イオン化効率 / トリアシルグリセロール

Outline of Final Research Achievements

A pretreatment method for subcellular 3D mass spectrometry imaging (MSI) was developed. Previously, by combining gas cluster ion beam (GCIB) with time-of-flight secondary ion mass spectrometry (TOF-SIMS), I could successfully reveal sub-nuclear chemical map with 3D-MSI. On the other hand, it was also revealed that subcellular lipid distribution cannot be maintained with fresh frozen fixation conventionally used for matrix-assisted laser desorption/ionization (MALDI)-MSI.To overcome this problem, fixation with minimal concetration glutardehyde was opted. By the minimal fixation, 3D distribution of triacylglycerol with palmitic acids and palmitoleic acids in lipid droplets of adipocytes was successfully visualized.

Free Research Field

分析化学

Academic Significance and Societal Importance of the Research Achievements

本研究課題によって開発した試料固定法は、培地組成をほとんど変えることなく室温で比較的短時間で行うことができる。毒性も低いことから通常の実験環境でも行えるため、培養条件や薬剤刺激、発生における特定の時期や細胞の質量分析イメージング解析や各種顕微鏡観察との比較を可能にするという学術的な意義をもつ。疾患や薬剤の効果などを分子組成の変化としてとらえるなど、安心安全な社会の実現へも貢献できると考えている。