Altelaar A F Maarten, Luxembourg Stefan L, McDonnell Liam A, Piersma Sander R, Heeren Ron M A
FOM Institute for Atomic and Molecular Physics, Kruislaan 407, 1098 SJ Amsterdam, The Netherlands.
Nat Protoc. 2007;2(5):1185-96. doi: 10.1038/nprot.2007.117.
Imaging mass spectrometry (IMS) allows the direct investigation of both the identity and the spatial distribution of the molecular content directly in tissue sections, single cells and many other biological surfaces. In this protocol, we present the steps required to retrieve the molecular information from tissue sections using matrix-enhanced (ME) and metal-assisted (MetA) secondary ion mass spectrometry (SIMS) as well as matrix-assisted laser desorption/ionization (MALDI) IMS. These techniques require specific sample preparation steps directed at optimal signal intensity with minimal redistribution or modification of the sample analytes. After careful sample preparation, different IMS methods offer a unique discovery tool in, for example, the investigation of (i) drug transport and uptake, (ii) biological processing steps and (iii) biomarker distributions. To extract the relevant information from the huge datasets produced by IMS, new bioinformatics approaches have been developed. The duration of the protocol is highly dependent on sample size and technique used, but on average takes approximately 5 h.
成像质谱（IMS）能够直接在组织切片、单细胞以及许多其他生物表面上对分子成分的身份和空间分布进行直接研究。在本方案中，我们介绍了使用基质增强（ME）和金属辅助（MetA）二次离子质谱（SIMS）以及基质辅助激光解吸/电离（MALDI）IMS从组织切片中获取分子信息所需的步骤。这些技术需要特定的样品制备步骤，以实现最佳信号强度，同时尽量减少样品分析物的重新分布或修饰。经过仔细的样品制备后，不同的IMS方法提供了一种独特的发现工具，例如用于研究（i）药物转运和摄取、（ii）生物加工步骤以及（iii）生物标志物分布。为了从IMS产生的海量数据集中提取相关信息，已经开发了新的生物信息学方法。该方案的持续时间高度依赖于样品大小和所使用的技术，但平均大约需要5小时。
