This test is similar to standard Fear Conditioning, where mice receive an aversive stimulus (US) paired to a tone (CS), however, in Trace Fear Conditioning the CS and US are separated by a short stimulus free period, thought to engage the hippocampus and prefrontal cortex. Mice learn to associate the CS with presentation of the US and exhibit a freezing response upon presentation of the CS.

Transgenic Alzheimer's mice (APP/PS1) are observed to freeze less compared to their Wild Type (WT) littermates upon presentation of the CS.