Studies are conducted in adult male and female Sprague Dawley rats (Envigo) in an AAALAC-accredited facility with approval from an Institutional Animal Care and Use Committee, and implemented in accordance with the Guide for the Care and Use of Laboratory Animals 8th Edition (National Research Council, 2011) with standards set by the National Institutes of Health.

Experimental procedure

The acetone evaporation test measures aversive behaviors triggered by the evaporative cooling of a drop of acetone placed on the plantar surface of the hind paw and is considered as a measure of cold allodynia (Choi et al., 1994).

The acetone evaporation test is performed on rats acclimated to individual chambers placed on a wire mesh floor. An acetone bubble is applied to the plantar surface of the hind paw, and animals are monitored for a response from 0-30 seconds following acetone application. To determine sensitivity to cold, the acetone is applied 5 times (5-minute intertrial interval) to each hind paw. The outcome measured is the total number of hind paw withdrawals per application. The experimental protocol varies depending on the pain model and whether the expected phenotype is unilateral (e.g., SNL) or bilateral (e.g., CIPN).


Choi Y, Yoon YW, Na HS, Kim SH, Chung JM (1994). Behavioral signs of ongoing pain and cold allodynia in a rat model of neuropathic pain. Pain, 59 (3): 369-376. PMID: 7708411 DOI: 10.1016/0304-3959(94)90023-X

National Research Council (2011). Guide for the Care and Use of Laboratory Animals: Eighth Edition. Washington, DC: The National Academies Press

This work was conducted by PsychoGenics Inc. (Paramus, NJ) in collaboration with PSPP, NINDS, NIH under contract # 75N95019D00026