Multiplexed detection and the establishment of a novel high-throughput method for human germ cell quality screening...

We report a rapid, sensitive, and high-throughput method for quality control of human sperm cells and oocytes staining based on the aggregation-induced emission feature of the tetraphenylethylene-based luminogen (TPE-Ph-In), which is mitochondria-specific. Germ cells are evaluated to assess fertility and to facilitate assisted reproduction. In regular clinical practice, sperm quality is determined on the basis of visual examination and mathematical models of the sperm cell number, motility, and morphology. The maturation of the oocyte is crucial for the developmental competence of the resulting embryo. Human in vitro fertilization (IVF) have indicated that delaying insemination improves fertilization rates, presumably by allowing the completion of cytoplasmic maturation for those oocytes that have not completely matured at the time. Therefore, a more reliable method to determine germ cell quality is needed. The mitochondrial membrane potential (MMP) of spermatozoa reflects the function and status of those cells. In oocytes, the distribution of mitochondria indicates the readiness of the cell for fertilization. Aggregation-induced emission luminogens (AIEgens) have good biocompatibility and photostability and produce low levels of background signal. There are about 100,000 mitochondria per fully-grown human oocyte. Mitochondria in mammalian oocytes are spherical with little cristae, supplying large scale of ATP for embryo development. Here, we expanded the use of TPE-Ph-Into determine germ cell quality on the basis of the MMP and the intracellular distribution of mitochondria. We stained clinical sperm samples from 36 patients with infertility, as well as four oocytes, with TPE-Ph-In and examined the cells by confocal microscopy and cell sorting analysis. Our results showed a positive correlation between the MMP and sperm cell motility, as well as the different distribution of mitochondria in oocyte. Thus, staining with TPE-Ph-In could be used to quickly determine germ cell quality in vivo, bringing new possibilities for applications of AIEgens in biomedical research and clinical trials.