Bodzon-Kulakowska Anna, Młodawska Wiesława, Mielczarek Przemyslaw, Lachowicz Dorota, Suder Piotr, Smoluch Marek
Department of Analytical Chemistry and Biochemistry, Faculty of Materials Science and Ceramics, AGH University of Science and Technology, A. Mickiewicza 30, 30-059 Krakow, Poland.
Department of Animal Reproduction, Anatomy and Genomics, University of Agriculture in Krakow, Al. Mickiewicza 24/28, 30-059 Krakow, Poland.
Materials (Basel). 2023 Feb 9;16(4):1479. doi: 10.3390/ma16041479.
Oocytes are a special kind of biological material. Here, the individual variability of a single cell is important. It means that the opportunity to obtain information about the lipid content from the analysis of a single cell is significant. In our study, we present a method for lipid analysis based on the MALDI-based mass spectrometry imaging (MSI) approach. Our attention was paid to the sample preparation optimization with the aid of a wet-interface matrix deposition system (matrix spraying). Technical considerations of the sample preparation process, such as the number of matrix layers and the position of the spraying nozzle during the matrix deposition, are presented in the article. Additionally, we checked if changing the 2,5-dihydroxybenzoic acid (DHB) and 9-Aminoacridine (9AA) matrix concentration and their solvent composition may improve the analysis. Moreover, the comparison of paraformaldehyde-fixed versus nonfixed cell analysis was performed. We hope that our approach will be helpful for those working on lipid analyses in extraordinary material such as a single oocyte. Our study may also offer clues for anybody interested in single-cell analysis with the aid of MALDI mass spectrometry imaging and the wet-interface matrix deposition method.
卵母细胞是一种特殊的生物材料。在此，单个细胞的个体变异性很重要。这意味着从单个细胞分析中获取脂质含量信息的机会很大。在我们的研究中，我们提出了一种基于基质辅助激光解吸电离质谱成像（MSI）方法的脂质分析方法。我们借助湿界面基质沉积系统（基质喷涂）对样品制备进行了优化。文章介绍了样品制备过程中的技术考量，如基质层数和基质沉积过程中喷嘴的位置。此外，我们还检查了改变2,5-二羟基苯甲酸（DHB）和9-氨基吖啶（9AA）基质浓度及其溶剂组成是否能改善分析。此外，还进行了多聚甲醛固定细胞与未固定细胞分析的比较。我们希望我们的方法将有助于那些从事诸如单个卵母细胞等特殊材料脂质分析的人员。我们的研究也可能为任何借助基质辅助激光解吸电离质谱成像和湿界面基质沉积方法进行单细胞分析的人提供线索。
