研究目的
To report the development of a new experimental MSI setup referred to as 'laser ablation-remote-electrospray ionization' (LARESI) and to present unique MS imaging results of human tissue, i.e. to demonstrate LARESI SRM targeted MS/MS imaging of frozen kidney tissue containing cancerous and non-cancerous regions.
研究成果
The study demonstrates the applicability of the novel ambient LARESI SRM/MRM MSI method for investigating and discovering cancer biomarkers in human tissue. The method offers a direct, pre-treatment-free approach to chemical imaging of complex biological samples, with potential for rapid and accurate detection of tumor markers.
研究不足
The study is limited by the small number of tissue samples and the focus on a specific set of metabolites. The comparison between the two MSI methods is preliminary and requires further validation with a larger sample size.
1:Experimental Design and Method Selection
The LARESI SRM MSI experimental setup was developed for imaging of target metabolites in human kidney cancer tissue. The method uses an IR laser for ablation and electrospray ionization for ion generation, coupled to a tandem mass spectrometer operated in SRM or MRM modes.
2:Sample Selection and Data Sources
Four tissue samples containing both cancerous and non-cancerous regions, obtained from three patients with renal cell carcinoma (RCC), were imaged. Sixteen endogenous metabolites were selected for analysis based on literature reports of their varying abundance between cancerous and non-cancerous areas.
3:List of Experimental Equipment and Materials
Nd/YAG-pumped, tunable OPO laser (IR Opolette 2731-HE; Opotek), SCIEX QTRAP 5500 mass spectrometer, silver-109 nanoparticles, stainless steel targets, HPLC quality solvents.
4:Experimental Procedures and Operational Workflow
Tissue sections were cut using a microtome and mounted on a Peltier stage set to -18°C. The LARESI SRM MSI and 109AgNPET LDI MSI methods were used for imaging. The spatial resolution was in a range from 175 to 300 μm.
5:Data Analysis Methods
MS images were analyzed for differences in abundances of selected metabolites between cancerous and non-cancerous regions. The data were processed using software described in the paper.
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