High spatial resolution imaging of biological tissues using nanospray desorption electrospray ionization mass spectrometry.,Nature Protocols

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High spatial resolution imaging of biological tissues using nanospray desorption electrospray ionization mass spectrometry.
Nature Protocols ( IF 13.1 ) Pub Date : 2019-11-13 , DOI: 10.1038/s41596-019-0237-4
Ruichuan Yin ₁ , Kristin E Burnum-Johnson ₂ , Xiaofei Sun ₃ , Sudhansu K Dey ₃ , Julia Laskin ₁

Affiliation

Mass spectrometry imaging (MSI) enables label-free spatial mapping of hundreds of biomolecules in tissue sections. This capability provides valuable information on tissue heterogeneity that is difficult to obtain using population-averaged assays. Despite substantial developments in both instrumentation and methodology, MSI of tissue samples at single-cell resolution remains challenging. Herein, we describe a protocol for robust imaging of tissue sections with a high (better than 10-μm) spatial resolution using nanospray desorption electrospray ionization (nano-DESI) mass spectrometry, an ambient ionization technique that does not require sample pretreatment before analysis. In this protocol, mouse uterine tissue is used as a model system to illustrate both the workflow and data obtained in these experiments. We provide a detailed description of the nano-DESI MSI platform, fabrication of the nano-DESI and shear force probes, shear force microscopy experiments, spectral acquisition, and data processing. A properly trained researcher (e.g., technician, graduate student, or postdoc) can complete all the steps from probe fabrication to data acquisition and processing within a single day. We also describe a new strategy for acquiring both positive- and negative-mode imaging data in the same experiment. This is achieved by alternating between positive and negative data acquisition modes during consecutive line scans. Using our imaging approach, hundreds of high-quality ion images were obtained from a single uterine section. This protocol enables sensitive and quantitative imaging of lipids and metabolites in heterogeneous tissue sections with high spatial resolution, which is critical to understanding biochemical processes occurring in biological tissues.

中文翻译：

使用纳米喷雾解吸电喷雾电离质谱法对生物组织进行高空间分辨率成像。

质谱成像 (MSI) 可以对组织切片中的数百个生物分子进行无标记空间映射。这种能力提供了关于组织异质性的有价值的信息，而这些信息是使用群体平均分析难以获得的。尽管仪器和方法都取得了重大进展，但单细胞分辨率的组织样本 MSI 仍然具有挑战性。在这里，我们描述了一种使用纳米喷雾解吸电喷雾电离 (nano-DESI) 质谱法对具有高（优于 10 微米）空间分辨率的组织切片进行稳健成像的协议，这是一种环境电离技术，在分析前不需要样品预处理。在该协议中，小鼠子宫组织被用作模型系统来说明这些实验中获得的工作流程和数据。我们详细描述了纳米 DESI MSI 平台、纳米 DESI 和剪切力探针的制造、剪切力显微镜实验、光谱采集和数据处理。受过适当培训的研究人员（例如，技术人员、研究生或博士后）可以在一天内完成从探针制造到数据采集和处理的所有步骤。我们还描述了在同一实验中获取正模式和负模式成像数据的新策略。这是通过在连续行扫描期间交替正负数据采集模式来实现的。使用我们的成像方法，从单个子宫切片中获得了数百张高质量的离子图像。

更新日期：2019-11-13

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