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Chemicals for assisted hatching
Michael L. Reed said on 30 October 2009
"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"
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Acid Tyrode solution
Thomas said on 26 October 2009
"Hi Swapnil,
Assisted hatching (AH)can be done using Acid tyrode's solution.
An embryo is stabilized with a holding pipette held at the 9 o'clock position and a 10 µm pipette containing acid Tyrode solution is expelled gently over a small are 15- 20µm at 3 o'clock position until the zona is dissolved.Aspirate all the expelled acid tyrode solution with the same pipette.After the procedure,the embryos are thoroughly washed in fresh callibrated Medium and cultured until the time of Transfer.You should able to finish this procedure within 5-7 seconds/Embryo.I hope this will help you.
THOMAS."
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