A multiple T-maze made of plexiglass was constructed to conduct spontaneous alternation testing. The initial alleyway was 86.9 cm in length and 16.5 cm in width, and crossed the first choice alleyway at 46.5 cm. The first choice alleyway was 96.9 cm in length and 17.6 cm in width. On either end of the first choice alleyway was a remaining second choice T-stem, which began equidistant from the initial alleyway. Each second choice T-stem was 62.2 cm in length and 16.5 cm in length. Plastic guillotine doors were inserted at each place that a choice was offered, as well as in the initial alleyway at 23.7 cm, for a total of ten doors.
Subjects were placed in the starting area in front of the first door for 10 s. Then the door was raised and subjects were free to roam the maze for 60 s. After subjects entered one of the choice arms the door was lowered to prevent it from going back into the arm. If no choice was made subjects were removed from the maze. Each subject underwent five trails consecutively with a 60 s intertrial interval. Each trail offered a maximum of two choices. Subjects were tested for spontaneous alternation.
The hypothesis was that control subjects would make an alternate choice, while experimental subjects would not alternate choices. For example, if a control subject chose to turn right on the first choice, then it should turn left on its second choice. However, if an experimental animal chose to turn right on the first choice, then it should turn right on its second choice also.