We all possess that urge to rise and climb the ladders of growth. Nevertheless, we have to start with the basics! Even if we are tempted to master IVF procedures such as ICSI or embryo transfer, we first need to be aware of the procedures which act as a foundation for them. Before dwelling into embryology which needs more responsibility and proficiency, Andrology provides us with that necessary base to begin our journey into IVF. Although IVF procedures are necessary to be done in the right way, ‘preparations’ is that invisible forte which is again overlooked conveniently.

Preparations can be broadly defined in many ways but mostly can be attributed to a day prior to ovum pick up and IVF/ICSI. This will include Media handling, preparing dishes, aliquoting or keeping the disposables ready. It can also be stretched a bit to include the other days too such as doing ET or freezing. Preparation can further include, ensuring proper storage and facilitating QA/QC and GLP!

Preparation especially attributed to aliquoting media and making the dishes falls on a trainee or a junior. It may seem an insignificant task but it carries a lot of weight as the embryos which later you create are going to be cultured in the dishes you make during preparations. A non-CO2 based media if we use by mistake to make the dishes and culture gametes inside a HERA incubator, it will result into a disastrous scenario! Whilst doing preparation a day before ICSI/IVF, we have to remember certain important steps. It is essential you ask the clinician about the number of follicles that are expected as the number of dishes you make and the amount of media you remove will all depend upon that number! A gap between the same can cause problems in case we get more follicles than the dishes we have prepared to accommodate them. Too many dishes also may result in wastage and adding to the cost factor as media is expensive. When you are making the dishes in the form of single drop culture or even group culture, ensure you immediately overlay media aliquoted in the dish with oil. Especially for single drop culture, I have seen people erroneously first making all the drops and then successively overlaying it with oil. Making drops for a plate should be followed with instantaneous addition of oil for that plate and then proceeding to the next one. This will help maintain the pH. The tubes should be aliquoted with media required for flushing or screening and should be tight lid as they will be harboring non- CO2 based media! These preparations should be done in the sterile flow of LAF with clean hands preferably wearing gloves. It is also better to calculate the amount of media required to aliquot into a tube and subsequently drops should be made. Making the drops directly from the bottle by immersing the pipette multiple times can cause unseen contamination. The person making the dishes should know which media is supposed to be applied on which day. Along with the media, disposables such as dishes, ICSI plates and other necessary components should be assembled inside the LAF.

Along with preparing dishes and media, preparation can also include the following activities. Care should be taken to check the expiry date of all the consumables and the media usage should be closely monitored. Expiry dates of filters in equipments should also be checked even if the clinic may have an AMC contract. It is also important to check if all the machines are properly working to avoid any unwanted surprises. Media when it is obtained from the vendor should be checked for its cold chain maintenance. Liquid nitrogen levels in the cryo tanks needs to be topped up and checking the stocks too is essential! These are some of those few activities which may include as a part of the preparation domain.

All said and done, you may still get people to do ICSI and monitor it in case you feel there are no results. But, preparations which is meant to be done by juniors is difficult to monitor owing to sloppiness, lack of enthusiasm in correcting their mistakes and taking it too much for granted as you feel other important procedures needs focus. A good culture system and good embryology skills need the support of good ‘preparations’ and its time we also shed a bit of light on it too!!!