Head birefringence properties are associated with acrosome reaction, sperm motility and morphology.

Birefringence in sperm heads reflects an organized and very compacted texture, indicating nuclear and acrosomal structural normality. This study performed a direct analysis of the acrosome integrity in single spermatozoa to verify whether a pattern of total or partial head birefringence reflected the acrosome status. The morphology in fresh samples was assessed according to World Health Organization criteria while the characteristics of birefringence were evaluated by polarized light. Acrosome integrity was evaluated by fluorescein isothiocyanate Pisum sativum agglutinin that binds selectively to the acrosome content. According to the results, a reacted acrosome was present in 96% of spermatozoa with partial birefringence and only in 35% of those with totally birefringent heads. A great proportion of sperm cells with normal morphology showed total birefringence both in the presence (59%) or in the absence of motility (45%; P < 0.01), while in morphologically abnormal spermatozoa the frequency of total birefringence was comparable to that of partial birefringence irrespective of motility (26% and 27%, respectively, in motile spermatozoa; 22% and 19%, respectively, in immotile spermatozoa). These data support a strong association between partial birefringence and reacted acrosome and show that the patterns of birefringence vary depending on sperm motility and morphology.

Prolonged absence of meiotic spindles by birefringence imaging negatively affects normal fertilization and embryo development.

Meiotic spindle (MS) assembly in human oocytes is a dynamic process that can be visualized by computer-assisted microscopy. At extrusion of the first polar body a spindle bridge is detected until the completion of telophase I and its reformation requires approximately 1h. This study analysed 396 oocytes from 112 cycles for fertilization and cleavage according to MS detection at two examinations, 39 and 41 h post-human chorionic gonadotrophin (HCG). All cycles had at least one injected oocyte lacking a visible MS at intracytoplasmic sperm injection (41 h post-HCG). To evaluate the results, oocytes were divided according to the presence (group A) or absence at both observations (group B) of the MS. Compared with group A, group B oocytes had lower normal fertilization rates, higher incidence of three pronuclei and two pronuclei in early dissolution and lower development to blastocyst. Some group A oocytes showed a late MS formation (not visualized at 39 h but at 41 h) and their performance was similar to that of the oocytes with a MS visible at both time points. Although some implantations occurred in group B, these findings suggest that prolonged MS non-detection could be a marker of reduced oocyte competence.

Impact of oocyte cryopreservation on embryo development.

OBJECTIVE: To verify whether the morphologic evaluation of zygotes and embryos derived from thawed oocytes could provide some relevant information regarding their developmental performance. DESIGN: Fertilization, zygote, and embryo morphology from sibling fresh and frozen oocytes was compared. SETTING: Reproductive Medicine Unit, Società Italiana Studi Medicina della Riproduzione, Bologna, Italy. PATIENT(S): Two hundred thirty-four patients underwent intracytoplasmic sperm injection cycles from which 1,101 spare metaphase II oocytes were cryopreserved. Subsequently, 256 thawing cycles were performed, and 997 oocytes were thawed. INTERVENTION(S): Intracytoplasmic sperm injection was performed on both fresh and frozen oocytes. MAIN OUTCOME MEASURE(S): Fertilization rates, pronuclear zygote morphology, and embryo cleavage rates. RESULT(S): Thawed oocytes had lower chances of being fertilized and developing into top-quality zygotes and regularly cleaving embryos when compared with sibling fresh oocytes irrespective of female age. As a result, the percentage of transferred cycles was significantly lower in frozen cycles compared with fresh cycles (79% and 93%, respectively); the proportion of transferred top-quality embryos followed the same trend. CONCLUSION(S): Reduced fertilization and cleavage rates in frozen cycles when compared with sibling fresh oocytes suggest that, even if surviving thawing, the process of slow freezing has a negative impact on the potential of further growth that is evident as early as the first cleavage divisions.