Micromanipulations are laboratory procedures that are used to assist in fertilization and implantation of embryos.

Intracytoplasmic sperm injection (ICSI):

it is the procedure when a single spermatozoon is injected directly in the egg. This procedure is done under an inverted microscope with magnification 200-400x. The holding pipette is 20 times thinner than human hair and the injection pipette is 60 times thinner than human hair.

Denudation of oocytes is an essential prerequisite for ICSI. The cells of the cumulus and corona cells are removed by combination of an enzymatic and mechanical procedure. The spermatozoon is immobilized and aspirated by the injection needle. The oocyte is fixed by holding needle and after that the spermatozoon is injected into cytoplasm of egg.

The injected egg is returned into incubator and 16-18 hours later is inspected by the embryologist for fertilization.

ICSI offers treatments for patients with impaired sperm function owing to:

• Oligozoospermia (less than 20 millions sperms);
• Asthenozoospermia (less than 30% motile sperms);
• Teratozoospermia (less than 5% normal forms, using the Kruger’s strict criteria);
• Low sperm survival;
• Patients with failure of fertilization in a previous IVF cycle;
• Patients with repeated IVF failed cycles.

In the last years ICSI’s use is extended to women with advancing age, women suffering from endometriosis and those with immunological and unexplained infertility.
In cases of Azoospermia, all the stages of spermatogenesis can be utilized for ICSI technique:

• MESA ( sperm retrieved from epididymis );
• TESA (sperm aspirated from testes);
• TESE (sperm extracted from testes).

Preimplantantation Genetic Diagnosis

This is a procedure that consists in isolation of one of the blastomers (the cells) of a growing embryo, which is subjected to genetic analysis for the purpose of finding genetic abnormalities and transfer (back) to the uterus of an embryo with normal genetic code.
The method is applied in cases with two or more spontaneous abortions, women in advanced age, multiple unsuccessful IVF procedures, chromosome translocations, and proven abnormal testicular meiosis of the man. The pre-implantation genetic diagnostics increases the conception rate and reduces the multiple pregnancy risk because of the small number of embryos transferred.

Embryo biopsy

Assisted Hatching

During the natural conception embryo hatching involves a spontaneous rupture of the zona pellucida (ZP), a membrane which surrounds the egg, to release the embryo for implantation into the womb. Sometimes, however, the zona pellucida is abnormally thick, preventing the embryo from hatching out. The failure of human IVF embryos to implant is thought to be partly due to the failure of their ZP to rupture. This means the embryo cannot implant into the womb and a miscarriage occurs. This medical problem is called 'implantation failure' and is more commonly experienced by older women, although it can happen in any age group. To help embryos to hatch from their ZP, assisted hatching procedures have been developed.

Assisted hatching is usually performed on embryos at the 6 to 8-cell stage just prior to embryo transfer). It has been suggested that the opening created in the zona assists hatching of the embryo and/or allows earlier interaction between the embryo and the endometrium thereby accelerating the implantation of embryos during this critical period of time.

The risks and discomforts of the clinical procedures are the same as for conventional IVF. There is a small risk of damage to the embryos during the hatching procedure.

However,to date there is no strong evidence strong evidence that is beneficial to use with all patients. It is recommended for women after 38 years of age, patients with reduced ovarian reserves, endometriosis and poor quality of the embryos. 

A fully hatched blastocyst with empty zona