期刊
INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES
卷 22, 期 19, 页码 -出版社
MDPI
DOI: 10.3390/ijms221910833
关键词
tissue sampling; tissue homogenization; nanosecond infrared laser; laser ablation; proteomics; mass spectrometry
资金
- Landesforschungsforderung (LFF) of the city of Hamburg [LFF-FV-75]
- Deutsche Forschungsgemeinschaft (DFG) [INST 337/15-1, INST 337/16-1, INST 152/837-1]
Recent studies have shown that nanosecond infrared lasers are effective for tissue sampling and homogenization, suitable for mass spectrometric proteomics research. Liquid chromatography tandem mass spectrometry analysis revealed significant abundance differences in proteins between two tissue types.
It was recently shown that ultrashort pulse infrared (IR) lasers, operating at the wavelength of the OH vibration stretching band of water, are highly efficient for sampling and homogenizing biological tissue. In this study we utilized a tunable nanosecond infrared laser (NIRL) for tissue sampling and homogenization with subsequent liquid chromatography tandem mass spectrometry (LC-MS/MS) analysis for mass spectrometric proteomics. For the first time, laser sampling was performed with murine spleen and colon tissue. An ablation volume of 1.1 x 1.1 x 0.4 mm(3) (approximately 0.5 mu L) was determined with optical coherence tomography (OCT). The results of bottom-up proteomics revealed proteins with significant abundance differences for both tissue types, which are in accordance with the corresponding data of the Human Protein Atlas. The results demonstrate that tissue sampling and homogenization of small tissue volumes less than 1 mu L for subsequent mass spectrometric proteomics is feasible with a NIRL.
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