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用于脂质组学研究的小鼠单卵母细胞基质辅助激光解吸电离飞行时间质谱成像

Mouse single oocyte imaging by MALDI-TOF MS for lipidomics.

作者信息

Bodzon-Kulakowska Anna, Arena Roberta, Mielczarek Przemyslaw, Hartman Kinga, Kozoł Paulina, Gibuła-Tarlowska Ewa, Wrobel Tomasz P, Gąsior Łukasz, Polański Zbigniew, Ptak Grazyna E, Suder Piotr

机构信息

Department of Biochemistry and Neurobiology, Faculty of Materials Science and Ceramics, AGH University of Science and Technology, Cracow, Poland.

Institute of Genetics and Animal Breeding, Polish Academy of Sciences, Jastrzebiec, Poland.

出版信息

Cytotechnology. 2020 Jun;72(3):455-468. doi: 10.1007/s10616-020-00393-9. Epub 2020 Apr 9.

Abstract

Reproductive cells are a very special kind of material for the analysis. Depending on the species, their dimensions allow for the application of mass spectrometry imaging-based techniques to receive a reasonable data for interpretation of their condition without any additional sample preparation steps, except for typical sample preparation characteristic for IMS protocols. A comparison between lipid profiles of oocytes could answer the question of the overall quality of the cells in the function of time or conditions of storage. Even tiny differences in the lipid profiles, but still detectable by bioinformatic analysis, could be crucial for the estimation of the conditions of the cells in various stages of development or aging. In our study, MALDI-TOF/TOF MSI was used to analyze and visualize the single oocytes. We deposited the cells on the transparent indium-tin-oxide (ITO) glass and marked their positions, which allowed for the fast localization of the cells and precise laser targeting in the ion source. We also optimized the usage of different MALDI matrices and different approaches. The proposed way of measurement allows analyzing quite a significant quantity of oocytes in a reasonably short time. During the analysis, the lipid composition of the single cell was successfully estimated in a conventional usage of the MALDI ion source, and the localization of lipids was confirmed by imaging mass spectrometry (IMS) analysis. The observed quantity of the lipids allowed for the application of the LIFT™ technique to obtain MS/MS spectra sufficient for lipids' unambiguous identification. We hope that our idea of the oocyte analysis will help to elucidate chemical changes that accompany different processes in which oocytes are involved. There could be such fascinating phenomena as the oocyte maturation, changes in the lipid components during their storage, and much more.

摘要

生殖细胞是用于分析的一种非常特殊的材料。根据物种的不同,其尺寸使得基于质谱成像的技术得以应用,从而在无需任何额外样品制备步骤的情况下获得用于解读其状态的合理数据,除了成像质谱(IMS)方案所特有的典型样品制备步骤。卵母细胞脂质谱之间的比较可以回答细胞在时间或储存条件作用下的整体质量问题。即使脂质谱存在微小差异,但仍可通过生物信息学分析检测到,这对于评估细胞在不同发育阶段或衰老阶段的状态可能至关重要。在我们的研究中,基质辅助激光解吸/电离飞行时间串联质谱(MALDI-TOF/TOF MSI)被用于分析和可视化单个卵母细胞。我们将细胞沉积在透明的氧化铟锡(ITO)玻璃上并标记其位置,这使得能够快速定位细胞并在离子源中进行精确的激光靶向。我们还优化了不同MALDI基质的使用和不同方法。所提出的测量方法能够在相当短的时间内分析相当数量的卵母细胞。在分析过程中,在MALDI离子源的常规使用中成功估算了单个细胞的脂质组成,并通过成像质谱(IMS)分析确认了脂质的定位。所观察到的脂质数量使得能够应用激光诱导热解吸(LIFT™)技术获得足以明确鉴定脂质的二级质谱(MS/MS)谱图。我们希望我们关于卵母细胞分析的想法将有助于阐明卵母细胞所涉及的不同过程中伴随的化学变化。可能会有诸如卵母细胞成熟、储存期间脂质成分的变化等迷人现象。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/2d27/7225219/782c069f4541/10616_2020_393_Fig1_HTML.jpg

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