Ambient mass spectrometry has been increasingly applied in tissue sample analysis with the advantages of high sensitivity,high speed and the capability for providing molecular structure information of compounds in complex matrices.Traditionally,the time-consuming sample pretreatment procedures including homogenization,extraction and separation are inevitable for tissue sample analysis,and commonly associated with biological degradation,chemicals contamination and bioactive components loss in sample preparations.Recently,with the development of novel ambient mass spectrometry techniques,in situ tissue samples analysis has become possible without sample pretreatment. The applications of ambient mass spectrometry in tissue samples analysis are mainly discussed in this review,and their impacts in different fields, such as life science,clinical medicine,food science and in vivo analysis are prospected.