The use of matrix-assisted laser desorption/ionization imaging mass spectrometry (MALDI-IMS) for the assessment of vascular pathology is attracting attention as a new valuable tool for diagnosing disease and finding new markers. MALDI-IMS is a molecular imaging technique whereby the simultaneous measurement of multiple samples directly from clinical tissue sections is possible. The versatility of MALDI-IMS has opened a new frontier in vascular pathology. In this review, we describe the principle and applications of MALDI-IMS.