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Dear IVF.net subscriber,
Welcome to the latest issue of IVF News.
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IVFMail
<< Add your questions here >>
| # |
Title |
Date Added |
|
Replies |
| 1 |
Blast Rate
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23 October 2009 |
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1
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| Dear Colleagues, please, would you be so kind to answer my question? How do you calculate the blastocyst rate: depending on number of fertilized eggs or number of embryos, developing on the 3th day and left up to 5-6 day of development?
Evgeny |
| 2 |
Zona thinning chemicals
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21 October 2009 |
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2
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| Is there any chemical to use for zona thining.
swapnil |
Replies
Re: Culture Dishes
Culture dishes
Antonia and Mike,
I am answering more on the section Mike wrote about using formalin in the AAB Proficiency Test of embryo medium. Yes, the adulterant is formalin and yes, without oil it can diffuse to other dishes; there was a paper on this a few years back now and I apologize for not remembering the authors name but it was in Fertil Steril and from the Cleveland Clinic showing the cross contaimination with formalin if oil not used.
To stop client complaints about not using a strong enough formalin concentration to get a well defined response of growth versus no growth of mouse embryos, human sperm motility and hamster sperm motility, the three major assays used in the AAB PT samples, we have to use a high concentration of formalin. Clinets and the AAB PT service miss the point entirely that what we are looking for in this PT is consistency between labs, not a growth/no growth situation. The AAB still fails to measure things such as Z-scores that would illustrate variability between labs for each particular sample. Sorry Antonia, got off track and on to my soap box; I will rereard your original post and comment if I can.
Patrick
Patrick Quinn Sage IVF
Re: Culture Dishes
Culture dishes
Antonia, me again. What media were being tested and what stage were the mouse embryos at initiation of culture, 1- or 2-cell? Do fresh mouse embryos work? I believe they can be obtained from Embryotech if you don't have a mouse colony. The reason I ask about stage and media is that I have found, as have others, that 1-cell embryos do not grow well in complex media such as Blastocyst medium, or probably KSOM with all amino acids (Life Global medium?) because the essential amino acids have a negative effect if growing from the 1-cell stage but not the 2-cell stage. Let's Mike, you and I have a brainstorming session at ASRM on this topic offer some strong coffees,
Patrick
Patrick Quinn Sage IVF
Re: Doubt (Statistics)
Doubt (Statistics)
I agree with Kim Pomeroy's comments here. Use the MedCalc package to construct Control Charts and you can then see if there are statistically significant variation in the parameters you are measuring over time.
Patrick
Patrick Quinn Sage IVF
Re: IVF Forum on the internet
IVF Forum on the internet
Linda, is your forum still functioning?
Patrick Quinn Sage IVF
Re: ph meter
Choice of pH meter
I would first echo what Patrick had said about reading up on pH, and add one reference to his selection, written by Rusty Poole. You can find the article at http://www.embryologists.com/acrobat/selected%20articles%20poolph.pdf.
Rusty also gives some advice on selection of pH probes. For myself, I've used an Oakton hand held unit for many years, with changing the probe every year. I now have one of the newer RI pH units, that uses much less medium and can used under oil, which is more reflective of how I do culture. The RI probe is a bit more complicated, but I like the results.
Mike
Michael L. Reed Center for Reproductive Medicine of New Mexico
Re: Slow development
Slow developing embryos
Dear Girón,
You did not mention about the state of embryo development in your laboratory before this problem was noticed.
Anyway, in any case, this is the time to do an intensive QC + QA exercise.
This quality control program starts from "me". I would see if my skills qualify to work in a clinical lab ? Whether I am following all the protocols religiously ? Then I would extend this to the laboratory to the OR room and to the transport system.
Go to the basics. Check whether all your equipment are working well. Incubators, microscope stages, tube warmers at correct temperature; incubator CO2 supply to give correct pH to your culture media; VOC level in your lab; quality of water in the incubators; temperature of the collection tubes at the time of oocyte retrieval, oocyte collection/flushing medium, the quality of air in the OR room, handling of gametes, temperature of the transporter cell; pressure of the suction pump etc.
There is another important factor which is often ignored. It is the skills of the clinician who is performing OR. Besides, stimulation protocols, quality of drugs, storage of the drugs, my God ! there are so many things.
In IVF, everything and every moment counts.
I know this is very cumbersome thing to do but there's no short cut. Please take help of some experienced person to troubleshoot the problem.
Sanjay Shukla
Re: Zona thinning chemicals
Acid Tyrode solution
Hi Swapnil,
Acid Tyrode can be used for Assisted haching.
Procedure: the micripippete containing acid tyrode's solution is brought very closely to the Zona pellucida and the acid solution is expelled gently over a small area (15-20 micrometer)until the Zona is dissolved.Aspirate all the expelled acid tyrode solution.This procedure you should not take more than 5-7 minutes.After the procedure, the embryos are throughly washed in a fresh callibrated medium and cultured until the time of transfer.
Acid tyrode is available in Medicult company.
I hope this will help you.
Thomas Madras Medical Mission
Re: Zona thinning chemicals
Chemicals for assisted hatching
Acidified Tyrode's solution is available from several vendors - this is the most common chemical method for assisted hatching. No need to make your own solutions anymore. I believe that the pH of acidified Tyrode's is around 2.0.
You could also investigate the use of pronase, although this has been used more for thinning the entire zona or removing the zona completely. There are a number of references on PubMed for this in non-human animal embryos as well as some for human embryos - you'll need to find the right concentration. Pronase has also been used to test the 'hardness' of the zona, by determining time from application of the enzyme to zona dissolution.
Mechanical methods include PZD using either a glass needle, or an opthalmic quality microblade. PZD methods require more technical skill.
Then there is laser ablation of the zona. Easier, but much more expensive.
Mike
Michael L. Reed Center for Reproductive Medicine of New Mexico
Re: Blast Rate
Calculating blastocyst development rate
I look at the data several ways, depending on the use of the data. If you are interested in total blastocyst rates, then calculating the rate from all fertilized ova is good - I believe that this measure is most often used for reporting in journals. I also break the data down further for cycles where embryos are replaced on day three or day five, as I believe that these cycles are very different in this practice. And if you are trying to predict development to day five, for example, using data from day three, then you could calculate blastocyst rates from the better quality embryos only. Periodically I'll identify three embryos on day three, that I would choose transfer if I were doing a day three transfer, and then track them to day five, to see if my choices were valid, to see how predictive my choices are, or are not.
With a spreadsheet or database program, you could do all of the above. Multiple metrics are good for troubleshooting.
Mike
Michael L. Reed Center for Reproductive Medicine of New Mexico
Re: Sticky eggs
hi dear.
i found the same big sticky cumulus surrounded the oocyte and when i tried to put in hyaluronidase become dark and adherent to oocyte but some oocyte i found was immature. also i am looking for the answer why this sticky cumulus and how i can managed. please answer me.
Dr alabayechi
alabayechi Kamal alsamaraee
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Data released today at the American Society for Reproductive Medicine Annual Meeting in Atlanta compares ART trends in the US with those around the world. Using data collected through the International Committee Monitoring ART (ICMART), investigators examined data from 2000- 2004. They found that delivery rates in the US were stable at 31% while they improved in Europe from 16% to 22%.
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