OBJECTIVE To evaluate changes in the dimension and volume of feline injection-site sarcomas (FISSs) before (in vivo) and after surgical excision and formalin fixation (ex vivo) as determined by measurements obtained from 2-D and 3-D CT images. SAMPLE 10 excised FISSs. PROCEDURES The maximum length, width, and depth of each FISS were measured on contrast-enhanced 2-D CT images of the tumor obtained in vivo and ex vivo. Those measurements were used to estimate tumor volume with the standard ellipsoid formula. Tumor volume was also calculated from 3-D CT images with software that used a volume-rendering algorithm. Student paired t tests were used for comparisons between the in vivo and ex vivo assessments. RESULTS Small decreases were detected in maximum tumor length, width, and depth between the in vivo and ex vivo assessments; however, tumor length was the only dimension that decreased significantly between the 2 assessments. Median tumor volume decreased significantly between the in vivo and ex vivo assessments regardless of the method used to estimate it. Tumor volume estimated by the ellipsoid formula was significantly lower than that estimated by the 3-D CT software at both assessments. CONCLUSIONS AND CLINICAL RELEVANCE Results indicated that shrinkage of FISSs following excision and formalin fixation was small and may be less than that of grossly normal tissue. Tumor volume estimated by the ellipsoid formula was consistently less than that estimated by 3-D CT software and should not be used when accuracy of tumor volume is of particular concern and advanced CT imaging is available.