At day 5 or 6 of culture embryos reach the blastocyst stage. At this time a laser-assisted biopsy is performed for pre-implantation genetic diagnosis. In other words, a very small piece of the trophectoderm is taken for analysis using a procedure capable of analyzing the entire chromosomes content (23 pairs). This technique allows patients and physicians to transfer embryos that are known to be genetically normal. The video shows a blastocysts hold into position and a biopsy micro-pipette applying suction while the laser is separating the cells for analysis at 400X magnification.

The in-vitro fertilization process is here compacted into a 26 seconds movie. The video starts with a mature oocyte showing its beautiful characteristics. Oocyte and corona radiate well-expanded right before insemination takes place. Oocyte is then inseminated, co-incubated overnight and fertilization is checked the next day showing 2 pronuclei. The embryo divided showing four and then eight cells ending at expanded blastocysts stage after culture for two more days.

The use of spermatozoa extracted from epididymis is required when there is a male factor due to obstruction or absence of the vas deference. In this case the urologists performs a small surgery extracting fluid for the visualization and preparation of spermatozoa for ICSI. In this video we show spermatozoa extracted from epididymis and placed in a culture dish where they swim and attach to a PICSI dish. The PICSI dish contains a substance (hyaluron), which is known to attach spermatozoa with high maturity and structural integrity. This procedure facilitates the spermatozoa selection for ICSI.

In some situations when the maturity and integrity of the spermatozoa requires confirmation, the laboratory performs the PICSI procedure. In this video it is shown how spermatozoa included in the sperm preparation are exposed to a plate containing hyaluron. Mature sperm are known to adhere to hyaluron while immature ones swim through without any adherence to the hyaluron spot. Before performing ICSI spermatozoa are loaded from the PICSI dish and placed into PVP before injection inside the oocyte.

In this video a greater detail of sperm immobilization is shown. As explained before this process is key for the normal fertilization of oocytes. Once the spermatozoon is immobilized it is loaded (tail-first) into the injection micropipette. Sometimes more than one spermatozoon is loaded to facilitate the injection process.

When ICSI is required, the motile spermatozoa have to be placed in a culture media containing a viscous substance called PVP. This media reduces the speed of the sperm so it can be immobilized and loaded into the injection micropipette. The immobilization process also activates the spermatozoa so it can perform a normal fertilization of the oocyte once it is placed in the oocyte cytoplasm.

In order to perform the insemination to oocytes the sperm has to be processed (washed). The process includes a filtration through density gradients with washes using culture media. From the total of spermatozoa included in a semen sample only a fraction is recovered. This fraction shows a very good motility (fast forward swimming) suitable for in-vitro fertilization or ICSI. In this video few spermatozoa are swimming across the counting chamber.

The physician in the operating room performs the oocyte aspiration (see video). The tubes obtained during that procedure are passed to the laboratory for oocyte identification. The process is performed in a sterile workstation using stereomicroscopes at 10-20X magnifications. As shown in the video the fluid contains several cells and in most cases the oocyte complex. This oocyte complex includes the oocyte (darker spot) the corona radiate and the cumulus oophorus. The oocyte complex is washed in culture medium and placed to culture in groups with sibling oocytes.

At day 2-3 or 5 of embryo culture embryos are transferred into the uterus. This process is performed under ultrasound guidance as shown in this video. The embryo/s are loaded into a catheter and the catheter introduced inside the uterus. Once the catheter is in place a very small amount of culture media is injected together with the embryo/s.