EMBRYOLOGY Lab Procedures: Step 1: Evaluation of Female Infertility
EMBRYOLOGY Lab Procedures: Step 1: Evaluation of Female Infertility
EMBRYOLOGY Lab Procedures: Step 1: Evaluation of Female Infertility
Background:
1. Female reproductive system
2. Oocyte Morphology
3. Buffers and Laboratory equipment knowledge
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● Step 3: Performing ICSI
○ The oocytes from the wash dish are transferred to the
HEPES drops in the ICSI dish after their denudation.
○ From the prepared semen, a drop from the top layer is
loaded to the PVP solution for sperm screening.
○ Before performing ICSI, neutralise the machine. Bring all
the controls to zero.
○ Attach the holding pipette and injecting pipette to their
respective holders.
○ Bringing them into position, use the injecting pipette to
aspirate out a morphologically normal sperm.
○ Use the holding pipette to keep the oocyte in place and
inject the single sperm into the oocytes with the help of
the injecting pipette.
● Step 4: Preparing culture dish
○ Use a 60mm dish.
○ Place three small CSC drops and one large CSC drop in
the dish.
○ Overlay with mineral oil.
○ Transfer the injected oocytes from the ICSI dish into the
first small drop of culture dish.
○ Keep washing the oocytes from the first small drop till
the third using flexi pets to completely be rid of the
MHM.
○ Place the oocytes in the large drop and incubate in dry
incubators for embryo day assessments.