Single Cell Profiling Using Ionic Liquid Matrix-Enhanced Secondary Ion Mass Spectrometry for Neuronal Cell Type Differentiation.
Single Cell Profiling Using Ionic Liquid Matrix-Enhanced Secondary Ion Mass Spectrometry for Neuronal Cell Type Differentiation.
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DOI：
10.1021/acs.analchem.6b04819
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发表时间：
2017-03-07
影响因子：
7.4
通讯作者：
Sweedler JV
中科院分区：
文献类型：
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作者：
Do TD;Comi TJ;Dunham SJ;Rubakhin SS;Sweedler JV
建立了一种用于基质增强二次离子质谱仪(ME-SIMS)的高通量单细胞图谱方法，用于研究神经细胞的脂质图谱。细胞群体分散在底物上，使用光学显微镜确定它们的位置，并使用细胞位置来指导从细胞获取SIMS光谱。一次实验可以检测多达2,000个细胞，每个细胞6个S。以离子液体为基质，从单个细胞中检测和验证了多种饱和和不饱和磷脂酰胆碱(PC)及其碎片。ME-SIMS光导单细胞图谱适用于各种大小的细胞，从大于75μm的加利福尼亚海兔神经元到7-μm的大鼠小脑神经元。ME-SIMS分析和在脂类分子质量范围(m/z 700-850)内峰的t分布随机相邻嵌入法区分了大鼠中枢神经系统的几种细胞类型，主要基于四种主要脂类的相对比例，PC(32：0)，PC(34：1)，PC(36：1)和PC(38：5)。此外，每种细胞类型内的亚群被初步划分为与其内源性脂质比率一致的亚群。这些结果说明了一种新方法的有效性，该方法使用SIMS对脂质和代谢物含量进行分析来对单细胞种群和亚群进行分类。这些方法广泛适用于高通量单细胞化学分析。
A high-throughput single cell profiling method has been developed for matrix-enhanced secondary ion mass spectrometry (ME-SIMS) to investigate the lipid profiles of neuronal cells. Populations of cells are dispersed onto the substrate, their locations determined using optical microscopy, and the cell locations used to guide the acquisition of SIMS spectra from the cells. Up to 2,000 cells can be assayed in one experiment at a rate of 6 s per cell. Multiple saturated and unsaturated phosphatidylcholines (PCs) and their fragments are detected and verified with tandem mass spectrometry from individual cells when ionic liquids are employed as a matrix. Optically guided single cell profiling with ME-SIMS is suitable for a range of cell sizes, from Aplysia californica neurons larger than 75 μm to 7-μm rat cerebellar neurons. ME-SIMS analysis followed by t-distributed stochastic neighbor embedding of peaks in the lipid molecular mass range (m/z 700–850) distinguishes several cell types from the rat central nervous system, largely based on the relative proportions of the four dominant lipids, PC(32:0), PC(34:1), PC(36:1), and PC(38:5). Furthermore, subpopulations within each cell type are tentatively classified consistent with their endogenous lipid ratios. The results illustrate the efficacy of a new approach to classify single cell populations and subpopulations using SIMS profiling of lipid and metabolite contents. These methods are broadly applicable for high throughput single cell chemical analyses.