Global Distribution Imaging, DMPK IM, AstraZeneca R&D, Södertälje, Sweden; Medical Mass Spectrometry, Department of Pharmaceutical Biosciences, Uppsala University, Uppsala, Sweden.
Medical Mass Spectrometry, Department of Pharmaceutical Biosciences, Uppsala University, Uppsala, Sweden.
J Proteomics. 2012 Aug 30;75(16):4912-4920. doi: 10.1016/j.jprot.2012.07.006. Epub 2012 Jul 14.
Analysis of whole animal tissue sections by MALDI MS imaging (MSI) requires effective sample collection and transfer methods to allow the highest quality of in situ analysis of small or hard to dissect tissues. We report on the use of double-sided adhesive conductive carbon tape during whole adult rat tissue sectioning of carboxymethyl cellulose (CMC) embedded animals, with samples mounted onto large format conductive glass and conductive plastic MALDI targets, enabling MSI analysis to be performed on both TOF and FT-ICR MALDI mass spectrometers. We show that mounting does not unduly affect small molecule MSI detection by analyzing tiotropium abundance and distribution in rat lung tissues, with direct on-tissue quantitation achieved. Significantly, we use the adhesive tape to provide support to embedded delicate heat-stabilized tissues, enabling sectioning and mounting to be performed that maintained tissue integrity on samples that had previously been impossible to adequately prepare section for MSI analysis. The mapping of larger peptidomic molecules was not hindered by tape mounting samples and we demonstrate this by mapping the distribution of PEP-19 in both native and heat-stabilized rat brains. Furthermore, we show that without heat stabilization PEP-19 degradation fragments can detected and identified directly by MALDI MSI analysis.
通过 MALDI MS 成像(MSI)对全动物组织切片进行分析需要有效的样本收集和转移方法,以允许对小组织或难以解剖的组织进行原位分析的最高质量。我们报告了在羧甲基纤维素(CMC)包埋动物的全成年大鼠组织切片过程中使用双面胶导电碳带的情况,将样品安装到大型导电玻璃和导电塑料 MALDI 靶上,从而能够在 TOF 和 FT-ICR MALDI 质谱仪上进行 MSI 分析。我们通过分析大鼠肺组织中噻托溴铵的丰度和分布情况,表明直接在组织上进行定量,表明安装不会对小分子 MSI 检测产生不当影响。重要的是,我们使用胶带为嵌入的脆弱热稳定组织提供支撑,从而能够对以前无法充分准备用于 MSI 分析的样品进行切片和安装,以保持组织完整性。通过对天然和热稳定的大鼠脑组织中 PEP-19 的分布进行映射,我们证明了这种方法不会妨碍较大肽组学分子的映射。此外,我们表明,如果没有热稳定化,PEP-19 降解片段可以直接通过 MALDI MSI 分析检测和识别。