Normative Preclinical Algesiometry Data on the von Frey and Radiant Heat Paw-Withdrawal Tests: An Analysis of Data from More Than 8,000 Mice Over 20 Years

The measurement of withdrawal to experimenter-delivered mechanical stimuli (von Frey test) and to heat stimuli (radiant heat paw-withdrawal or Hargreaves' test) applied to the hind paws is ubiquitous in preclinical pain research, but no normative values for the most-common applications of these tests have ever been published. We analyzed a retrospective data set of withdrawal thresholds or latencies in 8,150 mice in which these measures were taken using replicate determinations, before and after injection of inflammatory substances or experimental nerve damage producing pain hypersensitivity, totaling 97,332 measurements. All mice were tested in the same physical laboratory over a 20-year period using similar equipment and procedures. We nonetheless find evidence of large interindividual variability, affected by tester, genotype, mouse sex, tester sex, replicate order, and injury. These factors are discussed, and we believe that these normative data will serve as a useful reference for expected values in preclinical pain testing. PERSPECTIVE: This article presents a retrospective analysis of a large data set of mouse von Frey and radiant heat paw-withdrawal (Hargreaves' test) measurements collected in a single laboratory over 20 years. In addition to serving as a normative guide, sources of variability are identified including genotype, tester, and sex.