Abstract: 134

ERYTHROCYTE MONOLAYER ENABLES COMPUTER-ASSISTED MOTION ANALYSIS OF FOWL SPERM.

David Froman^{1 *}
Department of Animal Sciences, Oregon State University, Corvallis, Oregon ¹

Evaluation of sperm motility with the Hobson SpermTracker (Biogenics, Napa, CA) requires sample chambers of fixed depth, such as the MicroCell (Conception Technologies, San Diego, CA). In preliminary work, fowl sperm motility declined rapidly over an interval of minutes when sperm were evaluated at concentrations of 1.0 to 2.0 x 10⁶ sperm ml^–1 in MicroCells with a chamber depth of 50 µm. It was hypothesized that loss of sperm motility was due to either anoxia or oxygen toxicity. Therefore, a paired comparison was performed (n=14 roosters). Control sperm were diluted serially to 1.2 x 10⁶ sperm ml^-1 with 2 mM CaCl₂ in isotonic TES-buffered NaCl. Treated sperm were diluted similarly with the exception that the final dilution contained 3.5 x 10⁴ erythrocytes ml^-1. Sperm motion analysis was performed at 40°C using a 4x bright-field objective under a pseudo dark-field condition generated with a Ph3 annular phase ring. Sample chambers were filled with 7-µl volumes of sperm suspensions. When erythrocytes were present, these cells instantly formed a monolayer on the bottom of the chamber over which motile sperm moved. A fixed field of approximately 5 x 10⁸ µm² in the center of each MicroCell was evaluated over a 10-min interval. Repeated measurements (n=14) were made using 30-s sampling intervals with a 15-s pause in between. Mean linear velocity of control sperm decreased from 46 to 5 µms^-1. In contrast, the linear velocity of treated sperm was independent of time (P > 0.05). The Y-intercept for treated sperm was estimated to be 55 µms^-1 using the method of least squares. In summary, computer-assisted motion analysis of fowl sperm can be performed at extreme dilutions of sperm without artifact providing that the sample chamber contains an erythrocyte monolayer.

    This abstract is being presented on Sunday, August 1 at 8:00 AM to 10:15 AM at CUB 2nd Floor Ballroom.