High-Throughput Analysis of Tissue-Embedded Single Cells by Mass Spectrometry with Bimodal Imaging and Object Recognition
High-Throughput Analysis of Tissue-Embedded Single Cells by Mass Spectrometry with Bimodal Imaging and Object Recognition
复制标题
采用双峰成像和物体识别的质谱法对组织包埋的单细胞进行高通量分析
DOI：
10.1021/acs.analchem.1c00569
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发表时间：
2021
影响因子：
7.4
通讯作者：
Vertes, Akos
中科院分区：
文献类型：
--
作者：
Stopka, Sylwia A.;Wood, Ellen A.;Khattar, Rikkita;Agtuca, Beverly J.;Abdelmoula, Walid M.;Agar, Nathalie Y.;Stacey, Gary;Vertes, Akos
在生物组织中，细胞间的变异源于基因的随机和调节表达以及相应蛋白质的不同丰度。这些变异随后传播到下游代谢物产物并导致细胞异质性。质谱成像（MSI）是一种很有前途的工具，可以同时提供数百种生物分子的空间分布，而无需标记或染色。用于组织包埋单细胞直接分析的MSI仪器的技术进步主要是灵敏度、样品预处理和空间分辨率的提高，但受到低通量的限制。在此，我们介绍了一种双峰显微镜成像系统，该系统结合了基于光纤的激光烧蚀电喷雾电离（f-LAESI） MSI，并提高了组织嵌入单细胞（n> 1000）的吞吐量环境分析，以深入了解细胞异质性。基于自动图像分析，通过f-LAESI实现了精确的单细胞采样，从而发现了以不同代谢物水平为特征的细胞表型。
In biological tissues, cell-to-cell variations stem from the stochastic and modulated expression of genes and the varying abundances of corresponding proteins. These variations are then propagated to downstream metabolite products and result in cellular heterogeneity. Mass spectrometry imaging (MSI) is a promising tool to simultaneously provide spatial distributions for hundreds of biomolecules without the need for labels or stains. Technological advances in MSI instrumentation for the direct analysis of tissue-embedded single cells are dominated by improvements in sensitivity, sample pretreatment, and increased spatial resolution but are limited by low throughput. Herein, we introduce a bimodal microscopy imaging system combined with fiber-based laser ablation electrospray ionization (f-LAESI) MSI with improved throughput ambient analysis of tissue-embedded single cells (n> 1000) to provide insight into cellular heterogeneity. Based on automated image analysis, accurate single-cell sampling is achieved by f-LAESI leading to the discovery of cellular phenotypes characterized by differing metabolite levels.