Nagano Erika, Odake Kazuki, Shimma Shuichi
Research and Development Division, Miruion Corporation, Ibaraki, Osaka 567-0085, Japan.
Department of Biotechnology, Graduate School of Engineering, Osaka University, Suita, Osaka 565-0871, Japan.
Mass Spectrom (Tokyo). 2023;12(1):A0128. doi: 10.5702/massspectrometry.A0128. Epub 2023 Jul 25.
Mass spectrometry imaging (MSI) is a well-known method for the ionization of molecules on tissue sections and the visualization of their localization. Recently, different sample preparation methods and new instruments have been used for MSI, and different molecules are becoming visible. On the other hand, although several quantification methods (-MSI) have been proposed, there is still room for the development of a simplified procedure. Here, we have attempted to develop a reproducible and reliable quantification method using a calibration curve prepared from tissue debris of a frozen section of a sample when we trim the frozen blocks. We discuss the reproducibility of this method across different sample lots and the effect of the biological matrix (ion suppression) on our results. The quantitative performance was evaluated in terms of accuracy and relative standard deviation, and the reliability of the quantitative values obtained by matrix-assisted laser desorption/ionization-MSI was further evaluated by enzyme-linked immunosorbent assay (ELISA). Our -MSI method for the quantification of dopamine in mouse brain tissue was found to be highly linear, accurate, and precise. The quantitative values obtained by MSI were found to be highly comparable (>85% similarity) to the results obtained by ELISA from the same tissue extracts.
质谱成像(MSI)是一种用于使组织切片上的分子离子化并可视化其定位的知名方法。最近,不同的样品制备方法和新仪器已用于MSI,并且不同的分子变得可见。另一方面,尽管已经提出了几种定量方法(-MSI),但简化程序的开发仍有空间。在这里,我们尝试开发一种可重现且可靠的定量方法,该方法使用在修剪冷冻块时从样品冷冻切片的组织碎片制备的校准曲线。我们讨论了该方法在不同样品批次中的重现性以及生物基质(离子抑制)对我们结果的影响。根据准确性和相对标准偏差评估定量性能,并通过酶联免疫吸附测定(ELISA)进一步评估基质辅助激光解吸/电离-MSI获得的定量值的可靠性。我们用于定量小鼠脑组织中多巴胺的-MSI方法被发现具有高度线性、准确性和精确性。发现通过MSI获得的定量值与从相同组织提取物通过ELISA获得的结果具有高度可比性(>85%相似性)。