October’s Tip of the Month – Trouble-Shooting in the IVF Lab

Each month we will give a tip on how to improve your results, sourced either from the literature or our own research.

Help! Things are not working…

Trouble-shooting in the IVF lab

When your results don’t go to plan, what should you do?  There are a number of reasons why results fall away for no apparent reason.  It is frustrating, and believe me, everyone has experienced it.

The most sensible thing to do is carefully analyse what critical-to-less critical change has been made to the lab during a period. Therefore factors to consider when trouble-shooting include:

  • Calibration of the incubators (regular CO2 and temperature checks are necessary).
  • Batch of oil and plasticware/glassware which oil and media are stored or aliquoted into.
  • Introduction of volatile organic compounds, whether it be that the cleaner has changed their cleaning practice without first telling you, or changing of gas cylinders for similar reasons.
  • For media, we do not recommend aliquoting into smaller amounts, as every new container can have a residual contaminant.
  • Watch your refrigerator temperatures, especially at the back, as these can edge down to 0°C and cause freezing.  Freezing media to store is very detrimental to the stability of particular components that make up the media.
  • Equipment validation checks – including microscopes/warm stages, heating blocks and portable incubators.
  • Using different heparin sources can influence fertilization results.  The heparin levels required should be characterised for each bull using abattoir-derived cumulus-oocyte complexes.  There are lots of different heparins available, make sure your using an IVF-recommended one – we use DBL Heparin Sodium Injection BP (porcine mucous) 1mL vials 5000IU.

For more information on this month’s tip, or any trouble-shooting queries contact admin@artlabsolutions.com

September’s Tip of the Month – Grading Embryos for Transfer

Each month we will give a tip on how to improve your results, sourced either from the literature or our own research.

Grading Embryos for Transfer

Grading embryos to select which ones are suitable for transfer is a matter of experience at judging their quality.  Usually, in a busy commercial laboratory, grading is done without the aid of a high-quality inverted microscope.  The embryologist relies on what they see under a dissecting microscope.

Tips to assist grading are to (obviously) roll the embryo to change the perspective, but equally so, change the light angulation (by adjusting the mirror) to change the contrast.  Changing the contrast to assist in grading is an incredibly powerful tool, and the most experienced embryologists will do so regularly.  There are several resources describing MOET (in vivo) derived embryos. The most comprehensive is found in the International Embryo Technology Society’s manual (see www.iets.org for details).

However, there are fewer resources that assist training to judge IVF embryos.  Refer to; https://www.aeta.org/docs/Evaluation_of_in_vitro_produced_bovine_embryos.pdf, written by Jen Barfield from Colorado State University.

paper reference

As we have mentioned in May’s Tip of the Month (use of serum), IVF embryo production conditions vary and this can change the embryo’s appearance significantly.

New research is focussing on using high quality imaging systems and machine learning algorithms to remove much of the ‘expertise’ factor out of the equation.  We have been watching the research in this area and, especially for the IVF embryo, we can see that products will be available in 12-24 months from now that provide a high prediction capability for subsequent pregnancy.  We will keep you informed of these developments in the future.

For more information on this month’s tip, contact admin@artlabsolutions.com

ART Lab Solutions Media Batch #107 Excels in Bovine Embryo Assay

The bovine embryo assay was performed externally by an independent laboratory in Victoria, Australia. COC’s were aspirated from abattoir-derived ovaries and cultured in a serum free system using ART Lab Solutions complete bovine media suite.

Zygotes were then cultured in vitro until day 8 of embryo development. Cleavage and blastocyst rates were recorded.

Batch #107

Figure 1. Results presented as 66% viable blastocysts from pooled cleaved number of embryos.

For previous bovine embryo assay batch testing visit Quality Control Batch Tests or for more information about our testing procedures contact admin@artlabsolutions.com 

August’s Tip of the Month – Collection Medium

Each month we will give a tip on how to improve your results, sourced either from the literature or our own research.

What should you look for in a cumulus-oocyte complex (COC) collection medium (OPU)?

ART Lab Solutions does not manufacture a cumulus-oocyte complex (COC) collection medium, as there are several suitable substitutes.  In particular, media such as Vigro Complete Flush and EmCare plus an anticoagulant (heparin) are all perfectly suitable for OPU.  What is not going to help your COCs to give the best results is using simple buffered media such as Dulbecco’s PBS, or even worse, straight unbuffered saline.  We have a lot of data from both cow and mouse IVF that collection into energy substrate-free and unbuffered solutions can negatively impact oocyte quality within minutes.  Indeed, when it was recommended to change to a more complete media to a commercial group who were collecting with saline supplemented with a little FCS some years ago, their embryo production rates instantly improved.

For those who work with abattoir-derived cattle COCs, the best collection media of all is the follicular fluid that you aspirate.  Just keep your COCs in the aspirant until you’re ready to place into maturation media.  The natural meiotic inhibitors found in follicular fluid will work to prevent rapid germinal vesicle breakdown in the oocyte, providing a more synchronous meiotic progression during IVM.

Unfortunately, working with other species such as sheep, requires media with anti-coagulants (heparin) for aspiration.

For more information on this month’s tip, contact admin@artlabsolutions.com

ART Lab Solutions to Exhibit at 2018 Canadian Embryo Transfer Association (CETA/ACTE) & American Embryo Transfer Association (AETA)

ART Lab Solutions will be an exhibitor at the 2018 CETA/AETA Joint Convention in Montreal, Quebec, Canada.


Conference Details: 

Date: September 27-29, 2018

Where: Hotel Bonaventure Montreal, Quebec, Canada

Come past our booth and learn how our complete cattle IVF product range can improve the efficiency of breeding programs. We look forward to seeing you there!


The CETA/ACTE is the national body serving and representing the interests of its members and the embryo transfer industry. The Association is committed to excellence and promotion of embryo transfer technologies within Canada and abroad. The AETA provides a link between the science laboratory and field application, uniting organisations and individuals in the US who are engaged in the embryo transfer industry.

To learn more about the conference visit https://www.aeta.org/2018/ 

July’s Tip of the Month – Oil on Oil

Each month we will give a tip on how to improve your results, sourced either from the literature or our own research.

The Oil on Oil

Many IVF labs use micro drops under oil for embryo production, to protect the media from osmotic change and reduce the degree of pH change when transitioning to-and-from the incubator environment.  However, many embryologists would like to eliminate oil from their IVF systems; its messy and is a source of impurities that may negatively impact embryo development and quality.  So why is oil the preferred option to protect the media from osmotic and pH changes, especially when using with microliter micro drops of media?
Mineral oil is unique in its properties, because its hydrophobic (‘excludes water’) nature but allows gas to exchange between atmosphere and media
Oil is virtually impermeable to water vapour and even under 38-39 degrees temperature, there is undetectable osmotic change over the culture peiod.  Most plasticware used for IVF is either polystyrene or polycarbonate, and these are more porous to water vapour than mineral oil.  This goes unnoticed, as the amount lost is related to the surface area, of which the largest is the media-oil interface.  Surround media with nothing but plastic at a high surface-area to volume ratio will cause osmotic change over several days.
Silicon oil is also less hydrophobic and enables water vapour to be lost.
This hydrophobic property of mineral oil is combined with an impressive property for gas exchange. Although not as efficient as direct aqueous equilibration, gas exchange through mineral oil occurs readily, enabling sufficient gas exchange to maintain pH equilibria when using bicarbonate/CO2 buffering.
Contamination of oil is an issue. In particular, oil is susceptible to contamination by volatile organic compounds, hydroperoxides and other oil-breakdown contaminates, especially if not stored well over time.  Oil is best stored in glass, especially if aliquotting from a freshly purchased batch. Testing to identify individual contaminants is difficult, but these can be the cause of either poor or variable results if using unscreened oils. We recommend IVF laboratories source oil either from human IVF media companies or purchase specific mouse embryo assay tested oil.
ART Lab Solutions can facilitate your MEA testing requirements for oils and other laboratory consumables and products if required.
For more information on this month’s tip, contact admin@artlabsolutions.com

June’s Tip of the Month – Shelf Life & Storage of Media

Each month we will give a tip on how to improve your results, sourced either from the literature or our own research.

Shelf Life & Storage of Media

Embryo production media is a perishable product that requires cold storage.  Environmental factors that affect a media’s shelf life are light (radiation) and oxygen, along with the media’s composition.  The more ‘complex’ in formulation, the more likely that oxidative damage can occur over time.
When bottling media, one of the keys to prolonging shelf life is minimising the amount of air (oxygen) that is present.  Vacuum packing or inert gassing (we use nitrogen) are ways to prolong stored media immediately after manufacture.  But once a bottle is open and exposed to air again (even if still kept at 4 – 8C), how long will it last?  We have studied this extensively and found that our base medium has a 4-week usable life (serum-free), which is why we recommend using before this time.
But how long can you keep stored medium?  It will really depend on the breakdown rates of individual components, and this is dependent on composition.  This breakdown may be greatly accelerated once the medium is opened, if it is ‘aged’ from storing.  There is no doubt that “fresh is best”.
In a previous “Tip of the Month”, we discussed that serum (or serum substitutes) may be added to reduce the variability in results.  Adding serum will also help to “cover-up” some of the degradation occurring in media over time.  Personally, I am sceptical of claims of 12-month or longer stored media shelf-life in serum-free media.  I would want to see molecular fingerprinting of the stability of the media after that length of storing.
We are researching a new fingerprinting method that will allow us to provide that quality assurance.  In the meantime, we are confident of a 3-month shelf-life of our N2-gassed media.
For more information on this month’s tip, contact admin@artlabsolutions.com