Troubleshooting in the IVF lab
Managing an IVF lab is no task at hand. With every waking day, I only pray that I do not come across any unsolvable problem. Right from the paperwork to handling gametes one needs to be highly vigilant and especially in the clean rooms such as the IVF lab.
Handling stock solutions/culture media.
There are 2 types of media used for IVF. The CO2 dependant and CO2-independent. All media are to be used when at 37.0 degrees. The CO2-dependent media requires equilibration. The general practice is to aliquot the media necessary for the next day’s case and keep it in the CO2 incubator for overnight equilibration. What can go wrong?
You might forget to keep the media for overnight equilibration. In such cases, do not panic. The CO2-dependent media requires at least a minimum of 4 hours of equilibration (time taken for the formation of bicarbonate and maintaining correct pH). You can reach the lab early in the morning. However, research suggests a good 8 hours of equilibration. Either way, while troubleshooting, make sure that you have 4 to 5 hours of time prior to the start of the case. It’s always good practice to see the case charts for the next day which can avoid unnecessary tension. Expired media: Check the media consumables every week or write down the list of all the media with their corresponding expiry days.
Equipment in the IVF lab.
We’ve got to admit that the microscopes we use are super fancy. You know what else is fancy? The huge amount of money we have to pay for a small fix. More than getting your car fixed! That is why it is very important to keep the equipment in the lab calibrated and serviced regularly. And also to know the function of each part of your microscope and incubator and other equipments.
What can go wrong?
Loose parts with your microscopes. You should always always have an allenkey (can be placed behind the micromanipulator) to tighten screws, to change angles.
Pressure control with your holding and injecting pipette holder. You can remove the pipettes, screw them open and simply place your thumb on the opening, release the pressure through the control. Take it close to your ear and release your thumb, you will hear a small sound something similar to closing a pressure-filled pipe and releasing it. If not, there must be some bits and pieces of glass from the pipettes used. Take a thin wire of sorts and insert it through the back of the holder and push it deep inside. Similar to cleaning a bottle with a long brush. This way, anything stuck in, will come out.
Issues with the light source. My microscope with the micromanipulator has both Hoffman modulation optics as well as DIC. However, for the DIC a small filter (analyzer) at the base of the apparatus is placed as a movable slide. There was this one issue I encountered one day where the droplets on the dish splits light into VIBGYOR. Later I was told that the DIC filter when moved over normal Hoffman optics splits light from droplets on a plastic dish. DIC is used in case of IMSI and for IMSI a glass dish is used which prevents the splitting of light as it has better optical quality.
Sudden power shut down while doing ICSI. A senior embryologist told me one day that there was power cute while doing ICSI. He tackled the problem by using torch in the line of light source. I actually tried this (not while doing ICSI) with semen sample on a slide. You can actually see the specimen clearly. However, you will need the help of another person for this.
I will be writing more on this in the future as and when I tackle problems.
Get in touch with me if you’ve experienced such problems and we can discuss the different solutions for problems!