zona free oocyte will be most fragile, that is all you handle very gently while doing ICSI. Why you want to do zona-free ICSI, why not with Zona-intact? pl educate me.
Second what for you want to make lysate from human oocyte?
I do agree with your answer that's its very fragile to do and i have the experience sir but some time during the process of DENUDATION(80IU/ml Hyase ) some time few oocyte will lyse means it will become zona free , so i thought lets try ICSI with that and see what happens , its just out of curiosity .
But the question is that will it fertilize i have tried twice on two lysed oocyte but it didnt fertilize , and secondaly how to judge whether the lysed oocyte is from M 1 of M2?
OK that clarifies. I assume that "denudation time" under hyaluronidase treatment may vary from oocyte to oocyte (i.e. patient to patient, it needs subjective judgement to reduce the time of hyaluronidase (while watching under microscope) and immediate subsequent more washings to get rid of hyaluronidase,
Second, I can not comment on Lysate under M1 or M2,
I think it is unnecessary to make injections to oocytes without zona pellucida, it's just a loss of time. I experienced this few years before with this kind of oocyte and in the end no réultats were obtained. Otherwise there is nothing to differentiate the lysed oocyte at the stage M1 or M2.
It is consuming the extra time on the zona free oocyte . if you want to get experience ,its up to you.as I have experienced, after fertilization you will get a mass of cells that it does not able to progress the advanced development as blastocyste. if you decrease the pressur of holding you will do it