Automated motility and morphology measurement of live spermatozoa

Abstract

Background: Automated sperm analysis has wide applications in infertility diagnosis.Existing systems are not able to measure sperm count and both motility and morphology of individual live spermatozoa. Morphology measurement requires invasive staining, making the spermatozoa after morphology measurement not applicable to infertility treatment.Objective: To evaluate the reproducibility and reliability of automated measurement of individual live sperm's motility and morphology.Materials and methods: Fresh semen samples were obtained from twenty male partners attending for fertility investigations. The system firstly measured motility for all the spermatozoa within the field of view under a low magnification (20×), then a spermatozoa of interest is selected by the user and automatically relocated by the system after switching to a high magnification (100×) for morphology measurement.Reproducibility of sperm measurements was evaluated by intraclass correlation coefficients on consecutive measurement. Reliability of motility and morphology measurement was evaluated by tracking error rate and limits of agreement, respectively, with manual measurement as benchmark.Results: Measurement of all motility and morphology parameters had intraclass correlation coefficients higher than 0.94. Sperm motility measurement had a tracking error rate of 2.1%. Limit of agreement analysis indicated that automated measurement and manual measurement of sperm morphology were interchangeable. Automated measurement of all morphology parameters was not statistically different from manual measurement, as confirmed by the paired sample t test.Discussion: Automated motility and morphology measurement of single sperm revealed high reproducibility and reliability. The system also achieved a high efficiency for motility and morphology measurement. In addition to the intracytoplasmic sperm injection (ICSI) samples with polyvinylpyrrolidone (PVP), the developed sperm measurement technique is also effective for analyzing semen and washed samples. The system provides a valuable tool for quantitative measurement and selection of single spermatozoa for ICSI. It can also be used for sperm motility and morphology analysis in andrology laboratories.

Effects of six kinds of sperm staining methods on human sperm size and evaluation of their staining effects

“…4 Sperm morphological analysis is one of the critical indices for evaluating sperm quality. 5 Although there are reports on the morphological analysis of live sperm, 1,6,7 routine sperm morphological analysis is still based on the evaluation of stained sperm. In addition to Papanicolaou staining, Diff-Quik staining, and Shorr staining recommended by the World Health Organization (WHO), 8,9 Hematoxylineosin (HE) staining, Wright staining, and Wright-Giemsa staining are also used for sperm morphological analysis in some laboratories.…”