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Imaging Metals in Brain Tissue by Laser Ablation - Inductively Coupled Plasma - Mass Spectrometry LA-ICP-MS
Published on: January 22, 2017
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Infrared laser ablation sampling coupled with data independent high resolution UPLC-IM-MS/MS for tissue analysis
Michael E Pettit1, Fabrizio Donnarumma2, Kermit K Murray2
1Department of Chemistry and Biochemistry, Baylor University, Waco, TX, 76798, USA.
Analytica Chimica Acta
|September 9, 2018
Summary
Ion mobility-enhanced data-independent acquisition (HDMSE) mass spectrometry identified significantly more peptides and proteins from rat brain tissue compared to data-dependent acquisition (DDA). HDMSE offers superior proteomic analysis, even with challenging samples.
Area of Science:
- Proteomics
- Mass Spectrometry
- Neuroscience
Background:
- Bottom-up proteomics is crucial for understanding complex biological systems like the brain.
- Traditional mass spectrometry methods, such as data-dependent acquisition (DDA), can be limited by peptide ionization and detection efficiency.
- Optimizing sample preparation and data acquisition strategies is essential for comprehensive proteomic profiling.
Purpose of the Study:
- To compare the performance of ion mobility-enhanced data-independent acquisition (HDMSE) with data-dependent acquisition (DDA) for mass spectrometry-based proteomics of rat brain tissue.
- To assess the impact of ultra-performance liquid chromatography (UPLC) separation on the efficiency of both HDMSE and DDA.
- To evaluate the utility of infrared laser ablation microsampling in conjunction with these mass spectrometry techniques.
Main Methods:
- Rat brain tissue was analyzed using infrared laser ablation microsampling.
- Proteomic analysis was performed using mass spectrometry with both data-dependent acquisition (DDA) and ion mobility-enhanced data-independent acquisition (HDMSE).
- Whole tissue digests were analyzed across six different UPLC separation times to evaluate peak congestion effects.
Main Results:
- HDMSE consistently identified approximately seven times more peptides and four times more proteins than DDA from laser ablated samples.
- HDMSE analysis yielded about four times more identified proteins than DDA, irrespective of UPLC separation time for whole tissue digests.
- These findings highlight the superior depth of proteomic coverage offered by HDMSE.
Conclusions:
- HDMSE provides significantly enhanced proteomic coverage compared to DDA for rat brain tissue analysis.
- The benefits of HDMSE in protein identification are maintained even under conditions of UPLC peak congestion.
- Infrared laser ablation microsampling is a viable technique for preparing tissue samples for advanced proteomic analysis using HDMSE.
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