研究目的
To develop a method for direct analysis and two-dimensional mass spectrometry imaging of mouse bones without chemical treatments prior to analysis, preserving their physiological and chemical properties.
研究成果
The developed method allows for direct analysis and MSI of fresh, undecalcified mouse bones using IR-MALDESI, avoiding excessive sample preparation steps that modify the bone. This approach enables the detection of a broader range of endogenous biomolecules and demonstrates the feasibility of IR-MALDESI for 2D MSI of bones.
研究不足
The method is limited by the need for precise sample preparation to ensure uniform sampling and the challenge of analyzing bones without chemical modifications. The study also notes the difficulty in sectioning unmodified mouse bones due to their tendency to curl or fall apart.
1：Experimental Design and Method Selection
The study utilized IR-MALDESI MSI for direct analysis of mouse bones. The method involved flash-freezing bones, embedding them in Plaster of Paris, and imaging using IR-MALDESI coupled with a Q Exactive Plus mass spectrometer.
2：Sample Selection and Data Sources
Bones from healthy and stroke-affected C57Bl/6J mice were used. The bones were flash-frozen and embedded in Plaster of Paris for analysis.
3：List of Experimental Equipment and Materials
Q Exactive Plus mass spectrometer (Thermo Fisher Scientific),IR-MALDESI source,Plaster of Paris (DAP Products Inc.),Optima? LC/MS grade water (Fisher Scientific),Formic acid (Sigma-Aldrich),Methanol (Burdick and Jackson)
4：Experimental Procedures and Operational Workflow
Bones were flash-frozen, embedded in Plaster of Paris, and placed on the IR-MALDESI imaging stage. The stage was cooled, and the sample was equilibrated before imaging. A mid-IR laser pulse was used to desorb neutrals from each voxel on the bone, which were then ionized and analyzed by the mass spectrometer.
5：Data Analysis Methods
Data were processed in XCalibur, converted into .mzML format using MSConvertGUI, and then into .imzML files. Features were annotated using METASPACE, and ion heatmaps were generated using MSiReader.
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