Laboratory methods at the ISCARE clinic

ICSI - intracytoplasmic sperm injection is a micromanipulation technique that involves the direct injection of a single sperm into an egg (oocyte) using a special microscope.

This method is used when fertilisation fails or in the case of poor sperm quality (the number of sperm is insufficient, sperm motility is poor or the sperm are pathological). Sperm collected from the testis or epididymis using microsurgical techniques (MESA/TESE) may also be used in these cases.

PICSI (preselected intracytoplasmic sperm injection) is used to select the sperm that will be used for insertion into the oocyte. It is a so-called functional test.

A special gel is placed at the bottom of a Petri dish, which is used for this test. A medium containing sperm is subsequently added. The sperm migrate toward the gel and only mature sperm are capable of binding to the gel. The embryologist then uses these bound sperm for introduction into the oocyte (egg) using ICSI.

Cryoconservation or cryopreservation is a process aimed at preserving germ cells or embryos in a frozen state within liquid nitrogen for later use.

Cryoconservation uses media that contain cryoprotective agents- substances that protect cells during any fall in temperature. This is a safe method for freezing biological material.

Sperm cryoconservation (sperm banking)

In assisted reproduction, semen cryoconservation is a well-established and safe method that has been used successfully for a number of years now. The quality of the cryopreserved sperm is completely comparable with that of fresh sperm. The client´s own sperm, donor sperm or sperm collected using microsurgical procedures may be used.

Embryo cryoconservation

Embryo cryoconservation is an integral component of assisted reproduction. It predominantly serves to store surplus embryos (those not used for embryo transfer) in a given cycle. Only well-developed embryos can be frozen. In other cases, we freeze embryos for patients at a higher risk of hyperstimulation syndrome, those with insufficient uterine mucosa growth, and those with pathological findings on their reproductive organs that impede embryo implantation or in patients suffering from an acute illness that makes pregnancy impossible.

Oocyte cryoconservation (egg banking)

Oocyte cryoconservation is recommended for patients prior to any oncological treatment. Oocyte (egg) freezing is also promoted as so-called Social Freezing. This involves oocyte (egg) cryoconservation of women´s own germinal cells for a later time when they might want to decide to plan pregnancy.

Vitrification refers to the rapid cooling of cells. This is a method that is slowly beginning to replace standard cryoconservation (slow programmable freezing). It also represents the only option for freezing oocytes (eggs).

The classical cryoconservation method (slow programmable freezing) enables the freezing of embryos and sperm only.

Prolonged cultivation is a laboratory technique that enables the cultivation of embryos for more than 72 hours. We most frequently cultivate embryos for 4-5 days, preferably until the blastocyst stage.

In the five-day old embryo, it is possible to evaluate the formation of the embryo itself as well as of the embryonic layers. There is an increased chance of pregnancy following prolonged cultivation.

Semen analysis is an essential method for diagnosing male infertility. The basic indicators of sample quality include volume, appearance, total sperm count and motility.

Based on the semen analysis results, the physician sets out the therapeutic course to be followed.

You do not need to make an appointment for this examination. Semen analysis is performed from Monday to Friday, from 8:00 am to 3.00 pm. Once you have provided your sample, you may phone approx. 2 hours later for the result.

3-5 day sexual abstinence is necessary before sample collection. You may bring the sample from home (within 60 minutes of collection at body temperature) in a container that you can pick up at our reception on the 1st floor. Unfortunately, we cannot accept any other containers.

If you live far away, our clinic naturally provides a special collection room for men. On handing the sample over to the nurse, you must show some form of identification that includes a photograph (ID card, passport).

Definitions

EmbryoGlue increases the probability of embryo implantation within the uterus. The word “glue” in this term suggests that it works like a sort of bonding agent between the uterus and the embryo.

EmbryoGlue is enriched with carbohydrates, amino-acids and a high concentration of hyaluronic acid, all of which promote embryo implantation after transfer. With the help of these substances, a stronger bond between the embryo and the uterus is achieved and this minimises free movement of the embryo within the uterus.

EmbryoGlue is an option for all IVF patients undergoing transfer at our clinic.

Assisted hatching is a micromanipulation technique that involves erosion of the embryo´s external layer, thus facilitating the embryo´s implantation within the uterine mucosa.

AH is usually performed immediately before embryo transfer into the uterus, either mechanically using special needles attached to a manipulator or by laser.

MESA/TESE are microsurgical techniques used when there is a lack of sperm in the ejaculate (azoospermia), in erectile dysfunction or other similar disorders.

MESA involves aspiration of a cell suspension from the epididymis, which in some cases contains sperm. TESE involves the collection of tissue from which we subsequently attempt to acquire sperm.

The procedure is performed under general anaesthesia and lasts approximately one hour. After surgery, you will rest for 3 hours in our inpatient department and you may then leave, accompanied. Do not leave alone by public transport or car.

Before the procedure, you must undergo a preoperative evaluation at your GP´s, including an ECG. In our case, this exam is valid for three months. You will be provided with an order form by the nurses from the IVF centre on the 1st floor of the Clinic. On the day of your surgery, you should shave the area involved.

Good wound care and hygiene of the operated area must be insured after surgery. We also recommend that you refrain from wearing closely fitting underwear.

Pre-implantation diagnosis consists of a number of molecular-biological methods that aim to define the genetic status of the embryo.

Once the oocytes (eggs) have been harvested, fertilised and cultured (3 days), several cells are collected from suitable embryos. These cells are subsequently tested. The embryos are transferred on the fifth day.

If cells for analysis are collected from a five-day embryo, this embryo is not transferred but is vitrified (frozen).

PGD is used in cases where it is necessary to demonstrate or exclude a congenital defect of the foetus. Only healthy embryos are transferred or frozen for further use.

Pre-implantation genetic screening is a method that enables us to determine whether embryos resulting from IVF have not undergone random changes in their number of chromosomes.

This phenomenon occurs in nearly one half of embryos and is subsequently the cause of frequent miscarriages.

Cells are usually collected from five-day old embryos. The embryos are vitrified (frozen) and transferred during the patient´s subsequent cycle.

The most frequent chromosomal aberration during pregnancy is Down´s syndrome (trisomy of chromosome 21, whereby cells contain three chromosomes 21 instead of two).

We offer PGS e.g. to couples who have repeatedly undergone unsuccessful IVF or those with a history of recurrent miscarriages before the 12th week of pregnancy (women over the age of 35).

Continuous monitoring of embryos using the GERI device helps record and analyse the dynamics of embryonic development 24 hours a day.

The subsequent detailed analysis based on the acquired digital records enables an objective evaluation of the embryos.

The whole monitoring process takes place in a sealed incubator. One advantage is that it is not necessary to remove the embryos for the purpose of observation and the embryos are thus less exposed to unfavourable conditions during embryonic development. Besides the dynamics of embryonic development, the recording obtained can help assess embryo morphology in detail. We can thus select the best embryo for transfer. Moreover in some cases, we can decide, based on the recordings, when to potentially opt in a subsequent cycle for donor gametes (sexual reproductive cell).

In the case of certain methods (e.g. oocyte maturation under laboratory conditions in IVM), monitoring enables us to determine the ideal time for introducing the sperm into the oocyte (egg) via an injection (ICSI).

Sperm selection by MFSS chip

The method is used to select quality spermatozoa for later use in artificial insemination.

The microfluidic sperm selection chip ZYMOT (or FERTILE Plus) is a very gentle and chemical-free sperm processing method. The Zymot is a specially formed chamber with a selection membrane that mimics the natural environment of the female reproductive system (fallopian tubes). The selected spermatozoa have better morphology, genetic quality, viability and motility than the original sample.

The method thus represents a new and gentle alternative to standard sperm processing that does not require any preliminary treatment of the sperm sample and the use of devices (e.g. centrifuges) that increase the oxidative stress on the sperm. The sperm sample prepared in this way can be used for ova fertilisation by ICSI.

Note: the microfluidic sperm selection chip cannot be performed in cases where the sample does not contain a sufficient number of motile sperm. This method is not suitable for spermatozoa obtained by TESE and severe forms of oligoasthenoteratozoospermia.