Department of Veterinary Clinical Sciences , Small Animal Clinic, Institute of Veterinary Anatomy, Histology and Embryology, Justus Liebig University Giessen , Frankfurter Strasse 98 , 35392 Giessen , Germany.
IONTOF GmbH , Heisenbergstrasse 15 , 48149 Münster , Germany.
Anal Chem. 2018 Aug 7;90(15):8856-8864. doi: 10.1021/acs.analchem.8b00892. Epub 2018 Jul 9.
A method is described for high-resolution label-free molecular imaging of human bone tissue. To preserve the lipid content and the heterogeneous structure of osseous tissue, 4 μm thick human bone sections were prepared via cryoembedding and tape-assisted cryosectioning, circumventing the application of organic solvents and a decalcification step. A protocol for comparative mass spectrometry imaging (MSI) on the same section was established for initial analysis with time-of-flight secondary ion mass spectrometry (TOF-SIMS) at a lateral resolution of 10 μm to <500 nm, followed by atmospheric pressure scanning microprobe matrix-assisted laser desorption/ionization (AP-SMALDI) Orbitrap MSI at a lateral resolution of 10 μm. This procedure ultimately enabled MSI of lipids, providing the lateral localization of major lipid classes such as glycero-, glycerophospho-, and sphingolipids. Additionally, the applicability of the recently emerged Orbitrap-TOF-SIMS hybrid system was exemplarily examined and compared to the before-mentioned MSI methods.
一种用于高分辨率无标记人类骨组织分子成像的方法。为了保留脂质含量和骨组织的异质结构，通过冷冻包埋和胶带辅助冷冻切片制备了 4μm 厚的人类骨切片，避免了有机溶剂的应用和脱钙步骤。建立了一种用于同一切片的比较质谱成像 (MSI) 的方案，最初使用飞行时间二次离子质谱 (TOF-SIMS) 进行分析，横向分辨率为 10μm 至 <500nm，随后使用大气压扫描微探针基质辅助激光解吸/电离 (AP-SMALDI) Orbitrap MSI 进行分析，横向分辨率为 10μm。该方法最终实现了脂质的 MSI，提供了主要脂质类别的横向定位，如甘油酯、甘油磷酸酯和鞘脂。此外，还示例检查了最近出现的轨道阱-TOF-SIMS 混合系统的适用性，并与上述 MSI 方法进行了比较。
