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Poster Presentations
Day 4, June 25（Wed.）
Room P (Maesato East, Foyer, Ocean Wing)
- 4P-AM-16
Development of highly sensitive and comprehensive method for single-cell phospholipid analysis
(1Meijo Univ., 2Univ. Shizuoka, 3Yokogawa, 4ITO EN)
oTakuma Yanagisawa1,2, Eiji Sugiyama1, Yuta Terui3, Masafumi Iharada3, Hironori Takai3, Iwao Sakane4, Susumu Imanishi1, Kenichiro Todoroki2, Hajime Mizuno1
Phospholipids in mammalian cells play an essential role and have dynamics in various cellular functions; therefore, a quantitative analytical method for intracellular phospholipids is required. In our previous study, we developed an analytical method for single-cell phospholipids based on single-cell sampling with confocal microscopy followed by nanoESI-MS/MS. However, the number of detected phospholipid species was limited due to the low sensitivity of precursor ion scan used in that study. Herein, we have developed a highly sensitive analytical method by nanoESI-SRM, which enables quantitative and comprehensive evaluation of single-cell phospholipids. This method showed high sensitivity with an LLOQ at picomolar level and good linearity (R2>0.99). Furthermore, in measuring single-cell phospholipids of HepG2 cells, PCs and SMs were detected in positive ion mode. To obtain sufficient signal intensity from the larger number of phospholipid species, 10 cells were collected and concentrated as pooled samples prior to nanoESI-SRM. In addition to PCs and SMs detected in the single-cell sample, PEs, PSs, CLs, and PIs were detected from the 10-cell samples. The method developed in this study is expected to be used for monitoring dynamics at the single-cell level, in comprehensive phospholipid profiles linked to cellular functions.