A successful cryopreservation program has become the onus of fertility clinics all over. Importantly, as ‘freeze all’ cycles are slowly making inroads, the dependence on an effective cryopreservation program is the need of the hour.

Vitrification has been the choice lately for cryopreservation and can have slight variations depending upon the temperature, Cryo Devices and the brand of the media. Nevertheless, the basic principle and protocol are similar considering all these factors. Vitrification procedure is accomplished with the help of the available Vitrification media. The embryos are subjected to ES (Equilibration media) followed by its incorporation into VS (Vitrification media) before they are loaded on to a cryo device and plunged into liquid nitrogen. As an embryologist, I have always found Vitrification as more daunting to master than ICSI or other procedures. Unfortunately, there isn’t an exhaustive learning module with respect to the same. Fertility clinics do not follow a standard training program for Vitrification which may result into incompetency of the operator, poor survival rates and fluctuations in results!

The final step of embryo loading is the centerpiece of the whole procedure and the entire Vitrification program rests on the premise of that crucial last one minute. Naturally, the main focus will be on the last step which includes activities such as washing embryos in VS, loading them with minimal media on the cryo device and finally draining excessive media and plunging them into liquid nitrogen. The step is not difficult but certainly is challenging considering the microscopic aspects of the same coupled with the race with time.

The biggest challenge in Vitrification is the composite of speed with time! However, the technique can be effectively implemented after an extensive stepwise way to learn it by self devising a good training environment. The overall learning process may take longer to master but the initiation is critical.

Embryology Academy for Research and Training affiliated with Shivani Scientific Industries Pvt. Ltd. has designed a two-day ‘Advanced Course in Cryopreservation’ where beginning to learn Vitrification can be made easy. The course offers both theoretical aspects and hands-on experience with different cryo devices and even mimicking an actual freezing procedure. EART provides materials such as microbeads to practice the loading step which gives the trainee a hang of the technique and ushers in confidence. Training with respect to liquid nitrogen handling also is given along with brochures and study material. Tips such as placement of the hand whilst loading, the importance of labeling the devices, overcoming the fears of washing embryos in VS, the hand cryo device and eye coordination hurdles and steps associated with thawing are meticulously covered. Information is also given in terms of how the cryo devices will be stored and how the nitrogen tanks will be managed or maintained.

Nevertheless, after this critical initiation, if you get to work in a fertility clinic, there is an extensive way to further master the Vitrification procedure. The following are the steps over the course of six months you can chart in order to gain proficiency in the same!

Firstly, when you get acquainted with the technique just learning the protocol isn’t the key. There has to be an understanding of the science behind it. I have seen people panicking when they see embryos shrinking in ES!

Secondly, for starters, I would reckon an embryologist to be familiar with all the components of a Vitrification system. The goal here would be to do things perfectly without time constraints. The last major step can be broken into mini steps for better acclimatization. He can learn to adjust the coarse knob of the microscope to pick up the embryos after they float owing to change in viscosity in VS, the focusing of the cryotop under the microscope, the placement of the hand, learning the process of washing and finally gaining the ability to load the embryos!

Thirdly, once you have perfected the step, time will come into the picture. Then the use of stopwatch and practicing loading in its required time becomes the focus. Initially, the loading time may exceed even two mins. With practice, slowly, you will reduce the necessary time and finally place yourself in the stipulated range of sixty to ninety seconds! This practice can be done with degenerated oocytes.

Fourthly, once you have taken care of the time, the next natural factor will be survival! This is the stage where you will use discarded embryos and actually freeze them as if you are performing an actual procedure. Here, even the ES will come into your practice and you will observe intricate details such as the reaction of the embryos with the media. At this stage, the thawing technique will also be incorporated as it too contributes in the overall survival. The results can be recorded and it will give an insight into the overall improvement of the operator.

Finally, once you are done with survival, it is always recommended to freeze for embryo donation patients as ED cycle harbors the best quality embryos. Blastocysts should be considered more in the initial phase of freezing which survives better than say D2/D3 embryos. This can be furthered into increasing the number of cryo devices progressively corresponding with checking your performance of the previously frozen cryo devices simultaneously.

This comprehensive way of learning Vitrification may take six months overall depending upon the cycles happening in a fertility clinic which certainly affects the availability of gametes and resources(such as expired media for practice). It can help attain better proficiency over the technique and the program can be robustly standardized in a working fertility center itself. Vitrification is a gateway for the embryos to hibernate in an ice age and maybe we can make their journey qualitatively smooth!

And your journey with Vitrification can begin with Embryology Academy for Research and Training for a better know how and a better foundation!