Animals process and integrate sensory inputs to generate appropriate behavioral output. However, neural mechanisms underlying this process remain unclear. Here we established a short-term, visual discrimination task for head-fixed mice combined with in vivo two-photon imaging to relate neural activity to behavior. Viral vectors encoding the calcium indicator protein GCaMP6m were injected into the visual cortex of adult C57BL/6j mice. Water-restricted mice were trained to discriminate a target stimulus consisting of a grating drifting towards one horizontal direction, from a non-target stimulus drifting towards the opposite direction. The trained mice correctly responded to the target stimulus to lick a water-reward spout in front of the animal. After finishing the training, neural activity recording was carried out using two-photon calcium imaging from the mice performing the task. This head fixed, lick/no-lick visual discrimination task offers a quick learning paradigm compatible with pharmacological and neurophysiological methods including two-photon imaging, optogenetics and circuit tracing, making it possible to understand circuits and computations underlying perceptual decision-making as well as neurocognitive disorders.