A collective tracking method for preliminary sperm analysis

BACKGROUND: Total motile sperm count (TMSC) and curvilinear velocity (VCL) are two important parameters in preliminary semen analysis for male infertility. Traditionally, both parameters are evaluated manually by embryologists or automatically using an expensive computer-assisted sperm analysis (CASA) instrument. The latter applies a point-tracking method using an image processing technique to detect, recognize and classify each of the target objects, individually, which is complicated. However, as semen is dense, manual counting is exhausting while CASA suffers from severe overlapping and heavy computation. METHODS: We proposed a simple frame-differencing method that tracks motile sperms collectively and treats their overlapping with a statistical occupation probability without heavy computation. The proposed method leads to an overall image of all of the differential footprint trajectories (DFTs) of all motile sperms and thus the overall area of the DFTs in a real-time manner. Accordingly, a theoretical DFT model was also developed to formulate the overall DFT area of a group of moving beads as a function of time as well as the total number and average speed of the beads. Then, using the least square fitting method, we obtained the optimal values of the TMSC and the average VCL that yielded the best fit for the theoretical DFT area to the measured DFT area. RESULTS: The proposed method was used to evaluate the TMSC and the VCL of 20 semen samples. The maximum TMSC evaluated using the method is more than 980 sperms per video frame. The Pearson correlation coefficient (PCC) between the two series of TMSC obtained using the method and the CASA instrument is 0.946. The PCC between the two series of VCL obtained using the method and CASA is 0.771. As a consequence, the proposed method is as accurate as the CASA method in TMSC and VCL evaluations. CONCLUSION: In comparison with the individual point-tracking techniques, the collective DFT tracking method is relatively simple in computation without complicated image processing. Therefore, incorporating the proposed method into a cell phone equipped with a microscopic lens can facilitate the design of a simple sperm analyzer for clinical or household use without advance dilution.