An automated system for investigating sperm orientation in fluid flow

Abstract

Mammalian sperms reorient against fluid flow in the female reproductive tract, known as rheotaxis. Compared to chemotaxis that provides short-distance guidance, rheotaxis provides long-distance guidance for a sperm to find the egg cell. However, only a low number of sperms are capable of rheotaxis and their tail behavior during reorientation is not yet known. We have developed an automated system to manipulate human sperm orientation in fluid flow and quantitatively reveal sperm behavior changes during rheotaxis. The system automatically detects multiple sperms, selects the sperm for analysis, controls fluid flow, and quantifies sperm tail behavior. Sperm head angle is used as feedback to control fluid flow and select reorienting sperms. High accuracy of head angle tracking and automated sperm selection enables the capturing of dynamic sperm turning behavior in a large sample size. Algorithms are developed to track sperm tail skeletons and quantify tail beating amplitude and asymmetry, based on which the first quantitative analysis of sperm tail behavior in rheotaxis is obtained. Experimental results reveal, for the first time, that the sperms that are capable of reorienting against fluid flow beat their tails more asymmetrically than those sperms that are unable to reorient against fluid flow while no significant difference was found in their tail beating amplitudes.