The two-color miniscope imaging system use 470nm and 56l nm as excitation lights to record signals of both green and red fluorescence at the same site. When the reference channel uses the red fluorescent protein such as mCherry as a marker, the signal of the channel can be used as control data to exclude the motion noise (including the rotational noise of the fiber sliding ring) and verify the data validity of the calcium signal channel. The marker of the reference channel can also be replaced with a red calcium-sensitive fluorescent probe, and this system can simultaneously record the activity of two types of neurons in the relevant brain region in a certain behavioral paradigm, to reflect the encoding features of different types of neurons in the same behavioral paradigm.