Microdialysis is an excellent sampling technique for continuous monitoring of endogenous and exogenous compounds in the extracellular fluid. This technique has several advantages compared with conventional methods. Accordingly, this technique is extensively applied to study pharmacokinetics and pharmacodynamics both in experimental animals and human. In this mini-review, various topics related to microdialysis for the last five years will be discussed on its benefits. With the development of microdialysis technique and increasing number of applications of microdialysis for human study, this technique seems to play an important role in clinical practice, for example, bedside microdialysis as drug monitoring or diagnostic of disease.