Tricolor photography for assessment of spermatozoa motility

A rapid and accurate technique for assessment of spermatozoa motility has been developed. With the use of tricolor photography with sequential exposures in green, red, and blue light, spermatozoa were photographed in Makler chambers with a depth of 10 mu. Exposure in green light had a 1-second duration and was immediately followed by the red light from an electronic flash, and 0.05 second later by the blue light from a second flash. With this system, spermatozoa with motility would leave a green track, at the end of which they would appear first red and then blue. The colors and their intensity were chosen so that, when superimposed on nonmotile spermatozoa, the latter would appear white. By projection of the transparent film onto white paper, spermatozoa, motile and nonmotile, were easily counted and the percentage and motility was determined. Also, concentration could be calculated because the photograph represented a semen volume of 0.001 mu. Measurement of the green track and conversion to true length in micrometers gave the velocity in micrometers per second.