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Improved MALDI imaging MS analysis of phospholipids using graphene oxide as new matrix.

ABSTRACT: Matrix assisted laser desorption/ionization (MALDI) imaging mass spectrometry (IMS) is an increasingly important technique for detection and spatial localization of phospholipids on tissue. Due to the high abundance and being easy-to-ionize of phosphatidylcholine (PC), therefore, selecting matrix to yield signals of other lipids has become the most crucial factor for a successful MALDI-IMS analysis of phospholipids. Herein, graphene oxide (GO) was proposed as a new matrix to selectively enhance the detection of other types of phospholipids that are frequently suppressed by the presence of PC in positive mode. Compared to the commonly used matrix DHB, GO matrix significantly improved signal-to-noise ratios of phospholipids as a result of its high desorption/ionization efficiency for nonpolar compounds. Also, GO afforded homogeneous crystallizations with analytes due to its monolayer structure and good dispersion, resulting in better reproducibility of shot-to-shot (CV?

SUBMITTER: Wang Z

PROVIDER: S-EPMC5353552 | biostudies-literature | 2017 Mar

REPOSITORIES: biostudies-literature

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Improved MALDI imaging MS analysis of phospholipids using graphene oxide as new matrix.

Wang Zhongjie Z Cai Yan Y Wang Yi Y Zhou Xinwen X Zhang Ying Y Lu Haojie H

Scientific reports 20170315

Matrix assisted laser desorption/ionization (MALDI) imaging mass spectrometry (IMS) is an increasingly important technique for detection and spatial localization of phospholipids on tissue. Due to the high abundance and being easy-to-ionize of phosphatidylcholine (PC), therefore, selecting matrix to yield signals of other lipids has become the most crucial factor for a successful MALDI-IMS analysis of phospholipids. Herein, graphene oxide (GO) was proposed as a new matrix to selectively enhance ...[more]

PMID: 28294158

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Project description:Highly specialized cells are fundamental for proper functioning of complex organs. Variations in cell-type specific gene expression and protein composition have been linked to a variety of diseases. Although single cell technologies have emerged as valuable tools to address this cellular heterogeneity, a majority of these workflows lack sufficient in situ resolution for functional classification of cells and are associated with extremely long analysis time, especially when it comes to in situ proteomics. In addition, lack of understanding of single cell dynamics within their native environment limits our ability to explore the altered physiology in disease development. This limitation is particularly relevant in the mammalian brain, where different cell types perform unique functions and exhibit varying sensitivities to insults. The hippocampus, a brain region crucial for learning and memory, is of particular interest due to its obvious involvement in various neurological disorders. Here, we present a combination of experimental and data integration approaches for investigation of cellular heterogeneity and functional disposition within the mouse brain hippocampus using MALDI Imaging mass spectrometry (MALDI-IMS) and shotgun proteomics (LC-MS/MS) coupled with laser-capture microdissection (LCM) along with spatial transcriptomics. Within the dentate gyrus granule cells we identified two proteomically distinct cellular subpopulations that are characterized by a substantial number of discriminative proteins. These cellular clusters contribute to the overall functionality of the dentate gyrus by regulating redox homeostasis, mitochondrial organization, RNA processing, and microtubule organization. Importantly, most of the identified proteins matched their transcripts, verifying the in situ protein identification and supporting their functional analyses. By combining high-throughput spatial proteomics with transcriptomics, our approach enables reliable near-single-cell scale identification of proteins and profiling of inter-cellular heterogeneity within similar cell-types in tissues. This methodology has the potential to be applied to different biological conditions and tissues, providing a deeper understanding of cellular subpopulations in situ.

Dataset Information

Improved MALDI imaging MS analysis of phospholipids using graphene oxide as new matrix.

Publications

Improved MALDI imaging MS analysis of phospholipids using graphene oxide as new matrix.

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