Mass spectrometry imaging is an emerging label-free and molecular imaging technology. A complete and accurate mass spectrometry imaging data collection is inseparable from the pre-processing and post-processing operation. For the pre-processing steps, a complete pre-processing step should but not limited to include sample collection, sectioning, and sample fixation. The pretreatment steps should be imaginative, because it is not easy to make the sample into conductive slices. For the post-processing steps, a complete post-processing step should but not limited to include data processing, statistical analysis, and results discussion. To reflect pre-processing and post-processing operation more intuitively, use of video can make it easier for researchers to develop and promote operation procedures for mass spectrometry imaging. Researchers can use existing or self-developed new operation procedures and techniques. This is not only beneficial to the field of mass spectrometry imaging, but also common to other fields that require pre-processing and post-processing processing. In summary, this collection is dedicated to the development of pre/post-treatment techniques and methods related to mass spectrometry imaging and promotes them to serve a wider population of researchers.