Experimental and Data Analysis Considerations for Three-Dimensional Mass Spectrometry Imaging in Biomedical Research

Area of Science:

• Biomedical research
• Analytical chemistry
• Molecular imaging

Background:

• Mass spectrometry imaging (MSI) visualizes molecular distributions on surfaces.
• Current MSI studies primarily use 2D approaches, analyzing single tissue slices.
• Biological processes occur in 3D, necessitating volumetric analysis for comprehensive understanding.

Purpose of the Study:

• To review the expansion of MSI into three-dimensional (3D) analysis.
• To discuss experimental and data analysis considerations for 3D-MSI.
• To highlight developments enabling routine multi-sample 3D-MSI studies.

Main Methods:

• Expansion of Mass Spectrometry Imaging (MSI) techniques to acquire volumetric data.
• Development of data analysis strategies tailored for 3D molecular distribution datasets.
• Review of current advancements in 3D-MSI instrumentation and workflows.

Main Results:

• 3D-MSI provides visualization of molecular distributions within the entire sample volume.
• Investigating volumetric data offers a more comprehensive understanding of biological complexity.
• Single-sample 3D-MSI applications have rapidly increased.

Conclusions:

• 3D-MSI is crucial for fully understanding spatial and molecular complexity in biological samples.
• Addressing experimental and analytical challenges is key to successful 3D-MSI.
• Ongoing developments are paving the way for routine multi-sample 3D-MSI analysis.