Density gradient centrifugation is a common approach for preparing frozen-thawed semen for in vitro fertilization. This method is time and labor intensive, requires experience and sperm cell recovery is limited. Therefore, we tested a novel sperm purification device, the VetCountTM Harvester (MotilityCount ApS, Copenhagen, Denmark). The core elements of the VetCountTM Harvester are two chambers which are separated by a microporous membrane with a pore diameter of 10 µM. Spermatozoa are selected when actively swimming out of the semen-filled chamber, i.e., through the membrane, into the other chamber which contains a sperm collection medium. The handling of the VetCountTM Harvester is simple. Semen and collection medium, in this experimental approach a TALP (Tyrode’s Albumin Lactate Pyruvate) based medium, are injected into the chambers and, after 30 minutes incubation at 38°C, the medium is aspirated and the spermatozoa are ready for further use. In a preliminary evaluation, we assessed sperm recovery and semen quality of frozen-thawed but otherwise untreated bull semen and frozen-thawed semen treated with the VetCountTM Harvester or BoviPureTM gradient centrifugation (Nidacon International AB, Mölndal, Sweden), a standard technique in our laboratory. Frozen semen samples from six different bulls (n = 6), ten straws of one ejaculate per bull, were analyzed. Sperm concentration was determined using a hemocytometer chamber and the total sperm count was calculated. Motility parameters were assessed using IVOS II, a computer assisted sperm analysis (CASA) system, and flow cytometry was used to simultaneously evaluate viability, acrosome integrity, membrane fluidity and intracellular Ca+2 concentration. Results were tested for significant differences using Wilcoxon’s signed rank test with Bonferroni correction. A p-value of 0.05). The data demonstrate that the VetCountTM Harvester treatment selects a high-quality fraction of sperm from frozen-thawed bull semen with even lower free intracellular Ca2+ concentrations than a BoviPureTM gradient centrifugation. We are currently investigating whether sperm treated with a BoviPureTM gradient or the VetCountTM Harvester differ in cleavage rate, blastocyst rate and quality when they are used in bovine in vitro fertilization.