The startle response is an unconditional reflex, characterized by the rapid contraction of facial and skeletal muscles, to a sudden and intense startling stimulus. It is an especially useful tool in translational research for its consistency across species, simple neural circuitry, and sensitivity to a variety of experimental manipulations. The rodent acoustic startle response is commonly used to study fundamental properties of the central nervous system, including habituation, sensitization, classical conditioning, fear and anxiety, sensorimotor gating, and drug effects. The rodent startle response is typically assessed in stabilimeter chambers, and while these systems are excellent at measuring startle, they are designed only for this sole purpose. In the present study, we used the VideoFreeze system-a widely used tool for studying Pavlovian fear conditioning-to assess the acoustic startle response in freely moving mice. We validated the use of this system to quantify startle response amplitude and prepulse inhibition of startle. This is the first demonstration to date of using standard video in the automated assessment of the acoustic startle response in rodents. We believe that researchers already using the VideoFreeze system will benefit from the additional ability to assess startle without the purchase of new equipment.

The Pavlovian conditioned freezing paradigm has become a prominent mouse and rat model of learning and memory, as well as of pathological fear. Due to its efficiency, reproducibility and well-defined neurobiology, the paradigm has become widely adopted in large-scale genetic and pharmacological screens. However, one major shortcoming of the use of freezing behavior has been that it has required the use of tedious hand scoring, or a variety of proprietary automated methods that are often poorly validated or difficult to obtain and implement. Here we report an extensive validation of the Video Freeze system in mice, a "turn-key" all-inclusive system for fear conditioning in small animals. Using digital video and near-infrared lighting, the system achieved outstanding performance in scoring both freezing and movement. Given the large-scale adoption of the conditioned freezing paradigm, we encourage similar validation of other automated systems for scoring freezing, or other behaviors.