Experimental and Data Analysis Considerations for Three-Dimensional Mass Spectrometry Imaging in Biomedical Research
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REVIEW ARTICLE
Experimental and Data Analysis Considerations for Three-Dimensional Mass Spectrometry Imaging in Biomedical Research D. R. N. Vos,1 S. R. Ellis,1,2 B. Balluff,1 R. M. A. Heeren
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The Maastricht MultiModal Molecular Imaging Institute (M4I), Maastricht University, Universiteitssingel 50, 6229 ER, Maastricht, The Netherlands 2 Molecular Horizons and School of Chemistry and Molecular Bioscience, University of Wollongong, Wollongong, New South Wales, 2522, Australia
Abstract Mass spectrometry imaging (MSI) enables the visualization of molecular distributions on complex surfaces. It has been extensively used in the field of biomedical research to investigate healthy and diseased tissues. Most of the MSI studies are conducted in a 2D fashion where only a single slice of the full sample volume is investigated. However, biological processes occur within a tissue volume and would ideally be investigated as a whole to gain a more comprehensive understanding of the spatial and molecular complexity of biological samples such as tissues and cells. Mass spectrometry imaging has therefore been expanded to the 3D realm whereby molecular distributions within a 3D sample can be visualized. The benefit of investigating volumetric data has led to a quick rise in the application of single-sample 3D-MSI investigations. Several experimental and data analysis aspects need to be considered to perform successful 3D-MSI studies. In this review, we discuss these aspects as well as ongoing developments that enable 3D-MSI to be routinely applied to multi-sample studies. Key words: Mass spectrometry imaging, 3D imaging, Experimental set-up, Data analysis
Introduction Mass spectrometry imaging (MSI) is a label-free molecular imaging technique for which no prior knowledge about the sample is needed. It enables the visualization of molecular distributions on solid surfaces using mass spectrometry by scanning the samples in a pixel-by-pixel manner where for each pixel a mass spectrum is generated. Visualization of molecular spatial distributions provides unique insights in many fields such as material science and biomedical research [1, 2]. In the latter, such surfaces are typically thin planar sections prepared from biological tissue. MSI has
Correspondence to: R. Heeren; e-mail: [email protected]
been used extensively to image the two-dimensional (2D) distributions of endogenous or exogenous (drugs and pharmaceuticals) compounds in such planar tissue sections for the study of tissue-based diseases, tissue pharmacokinetics, or the study of biomaterials in medical applications [3, 4]. However, the natural state of the original sample is volumetric, hence keeping the original three-dimensional (3D) information is important to be able to study the structural context of the sample in all dimensions. In material science, where MSI has been employed since the 1960s [5], it is proven that 3D-MSI can provide essential information. Secondary ion mass spectrometry (SIMS), for instance, has been used extensively for 3D e
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