Lipidomics,firstly proposed in 2003,is an emerging field, where the structures, functions and dynamic changes of lipids in cells,tissues or body fluids are investigated.Studies have showed that the disturbance of lipid metabolism is closely related to some diseases,such as cancer,atherosclerosis,diabetes,obesity and Alzheimer disease.Therefore, the comprehensive analysis of lipid becomes significant. However, due to the diversity and complexity of lipids,lipids analysis is still full of challenges.Recently,Matrix-assisted laser desorption and ionization mass spectrometry imaging(MALDI MSI) has been developed quickly in lipidomics research. Compared with traditional LC-MS,MALDI MSI provides spatially resolved ion intensity maps corresponding to the mass-to-charge ratio of intact molecular species at specific locations in a prepared tissue sample, which is very useful in biomarker identification analysis,drug distribution analysis,etc. MALDI MSI is a relatively new technique,it still has many aspects needing further improved,such as small molecules detection, neutral lipids detection,ion suppression elimination,detection sensitivity improvement,etc.In MALDI MSI analysis,sample preparation,new matrix development and application research are the three hotspots.This review aims to introduce the research progress of MALDI MSI in sample processing,matrix coating and its application.The existing problems and possible solutions are discussed,in order to extend the application of MALDI MSI.