Micro-volume liquid handling technologies have been extensively used in the field of molecular biology and have been under rapid development over the last several decades. Accompanying the trend of laboratory automation and high-throughput, the liquid dispensing is increasingly playing an important role in protein crystallization, drug screening and DNA sequencing, and it also needs to satisfy the exacting requirements, such as the higher processing speed, higher pipetting precision and lower processing costs. The purpose of this review is to provide an overview of micro-volume liquid-handling technology and summarize the latest applications of this technology. Finally, some future trends and perspectives in these research areas will be outlined.