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This course will discuss the basics of data manipulation, visualization, and statistical analysis of mass spectrometry (MS) imaging experiments using Cardinal and R. We begin by introducing the fundamental ways of interacting with MS imaging datasets and experimental metadata using Cardinal and R, followed by tutorials on pre-processing and visualization of mass spectra and ion images. Finally, we will demonstrate the application and interpretation of statistical methods including segmentation, classification, and testing. The course is intended for scientists who are familiar with MS imaging (including DESI, MALDI, etc.), but who have limited exposure to programming with R. Chapters: 0:00:00 (Introduction) 0:08:08 (Cardinal 3 overview) 0:24:51 (Cardinal 3.6 changes and new data structures) 1:15:16 (Visualization with Cardinal 3.6) 2:32:00 (Summarization and preprocessing of MSI) 4:03:00 (Statistical analysis: segmentation) 4:59:10 (Statistical analysis: class comparison) 5:09:41 (Overview of coregistration of MSI and H&E images) 5:26:16 (Statistical analysis: classification (no audio)) Important resources relevant for the program: Dr. Kylie Bemis bio: https://www.khoury.northeastern.edu/p... Official Cardinal website: https://cardinalmsi.org Cardinal's Bioconductor page: https://www.bioconductor.org/packages... Cardinal GitHub repository: https://github.com/kuwisdelu/Cardinal Cardinal 3.6 guide: https://www.bioconductor.org/packages... Additional resources for the program: https://docs.google.com/document/d/1p... NOTE: The final 20 minutes did not capture the audio track, so it is without sound.