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DNA contamination from cumulus cells surrounding the egg or from unsuccessful sperm can contaminate PGD samples and result in misdiagnosis.
Fertilisation with ICSI involves careful removal of the cumulus cells followed by injection of a single sperm, thereby greatly reducing the risk of contamination.
Fertilised eggs are identified by the presence of pronuclei on day 1.
On day 3 of culture, the envelope enclosing the embryo (zona pellucida) is breached using a low power laser.
Zona breaching promotes the “hatching” of blastocysts on days 5-6, a little earlier than would occur with IVF.
A near infra-red laser creates a small slit (breach) in the zona pellucida.
The embryo grows to form a blastocyst - a clump of cells (inner cell mass) inside a hollow ball of trophectoderm cells.
The expanding blastocyst (now 120-150 cells) hatches through the small hole that was created in the zona on day 3.
Hatching blastocysts are held with gentle suction, then a biopsy pipette is used to remove 3-6 trophectoderm cells usually with the assistance of the laser.
The biopsied trophectoderm cells are ready for testing. The rest of the embryo is frozen to allow time for CGH analysis.
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