Influence of image sampling frequency on the perceived movement characteristics of progressively motile human spermatozoa.

Tracks of 30 progressively motile washed human spermatozoa were plotted manually from 200-Hz frame rate video recordings. Tracks at 100, 66.7, 50, 40, 33.3, 25, 20, 10, and 5 Hz were then constructed using every 2nd, 3rd, 4th, 5th, 6th, 8th, 10th, 20th, or 40th point. The 200-Hz tracks were analyzed by traditional manual methods, and all ten sets of tracks analyzed using a computer-assisted method ("Videomot," developed originally to analyze 30-Hz tracks) to eliminate observer bias. Progression velocity (VSL) remained constant under all analysis conditions. Average path velocity (VAP) also remained essentially constant, although Videomot was less reliable at high frame rates due to problems in determining the average path. Curvilinear velocity (VCL) was very frame rate dependent (the 25-Hz mean value was only 56.5% of that at 200 Hz), and Videomot was more accurate than manual analysis at 200 Hz. Values of the amplitude of lateral head displacement (ALH) were acceptable at most frame rates. At less than 66.7 Hz the inclusion of spurious curvilinear track deviations caused lower mean ALH values, and at 5 Hz ALH could not be measured since the track was essentially the average path. Beat/cross frequency (BCF) was also frame rate dependent; at high rates there was the same problem as with ALH measurements, while at less than or equal to 25 Hz the maximum BCF was restricted by the frame rate. We conclude that human sperm movement characteristics can be measured at frame rates ca. 30 Hz but only if the constraints affecting VCL and BCF values are understood and accepted. Finally, less than 10 Hz can only give values for VSL and, perhaps, VAP.