The acquisition of physiological signals is the first step of its analysis and processing. The development mode for PC/104 module is introduced and the design of multichannel acquisition system for physiological signals based on PC/104 is also put forward. The system hardware mainly includes PC/104 embedded computer,DIAMOND-MM-16-AT acquisition card and four-channel amplifiers. Windows 2000 is used as operating system for PC/104 embedded computer. The multithread program to sample data is made by means of programming language Visual C++ 6.0. The experimental results show that the system can be applied to sample, analyse, and process the physiological signals. The system also has lots of advantages, such as small volume, low power, firm structure and steady performance.