title: Study of Phospholipids in Single Cells Using an Integrated Microfluidic Device Combined with Matrix-Assisted Laser Desorption/Ionization Mass Spectrometry.
abstract: Single-cell trapping and high-throughput mass spectrometry analysis remain challenging now. Current technologies for single-cell analysis have several limitations, such as throughput, space resolution, and multicomponent analysis. In this study, we demonstrate, for the first time, the combination of microfluidic chip and matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS) for high-throughput and automatic single-cell phospholipids analysis. A microwell-array-based microfluidic chip was designed and fabricated for cell array formation on an indium tin oxide (ITO)-coated glass slide. Mass spectrometry imaging measurement with 25 μm pixel size was performed with a MALDI ion source. Eight phospholipids in a single A549 cell were detected, and their structures were further identified by MS/MS spectra. Selected ion images were generated with a bin width of Δm/z ± 0.005. The selected ion images and optical images of the cell array showed excellent correlation, and mass spectrometry information on phospholipids from 1-3 cells was extracted automatically by selecting pixels with the same fixed interval between microwells on the chip. The measurement and data extraction could be processed in several minutes to achieve a high-throughput analysis. Through the optimization of different microwell sizes and different matrices, this method showed potential for the analysis of other metabolites or metabolic changes at the single-cell level.
authors.affiliation: †Department of Chemistry, Tsinghua University, Beijing 100084, China.|‡State Key Laboratory Breeding Base-Shenzhen Key Laboratory of Chemical Biology, Graduate School at Shenzhen, Tsinghua University, Shenzhen 518055, China.
authors.forename: Weiyi
authors.initials: W
authors.lastname: Xie
authors.affiliation: ‡State Key Laboratory Breeding Base-Shenzhen Key Laboratory of Chemical Biology, Graduate School at Shenzhen, Tsinghua University, Shenzhen 518055, China.|§Key Laboratory of Metabolomics at Shenzhen, Shenzhen 518055, China.
authors.forename: Dan
authors.initials: D
authors.lastname: Gao
authors.affiliation: ¶Neptunus Pharmaceutical Technology Center, Shenzhen 518057, China.
authors.forename: Feng
authors.initials: F
authors.lastname: Jin
authors.affiliation: ‡State Key Laboratory Breeding Base-Shenzhen Key Laboratory of Chemical Biology, Graduate School at Shenzhen, Tsinghua University, Shenzhen 518055, China.|∇School of Medicine, Tsinghua University, Beijing 100084, China.
authors.forename: Yuyang
authors.initials: Y
authors.lastname: Jiang
authors.affiliation: ‡State Key Laboratory Breeding Base-Shenzhen Key Laboratory of Chemical Biology, Graduate School at Shenzhen, Tsinghua University, Shenzhen 518055, China.|§Key Laboratory of Metabolomics at Shenzhen, Shenzhen 518055, China.
authors.forename: Hongxia
authors.initials: H
authors.lastname: Liu
mesh_terms: D045744:Cell Line, Tumor; D006801:Humans; D046210:Microfluidic Analytical Techniques; D010743:Phospholipids / Q000032:analysis*; D059010:Single-Cell Analysis; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D017971:Tin Compounds / Q000737:chemistry
chemical_list: D010743:Phospholipids; D017971:Tin Compounds; C109984:indium tin oxide