Collection of Primordial Germ Cells from Drosophila Donor Embryos

Proceed to collect the primordial germ cells or PGCs from the dross filia embryos. After assembling the micro manipulator system. While working under a stereo microscope, use forceps to transfer the embryos.

Align the coated embryos in two rows on a PGC collection glass slide along two reference lines. Orient the embryos with their anterior to the right and ventral side up. Gently move the transplantation needle tip to the anterior end of an embryo.

Penetrate the embryo by moving the microscope stage and retract the needle slightly. When the tip reaches the posterior end. Discharge any yoke in the needle inside the somatic cell layer.

Move the needle tip to the posterior pole and gently load the PGCs into the needle without taking much time. Pull the needle out of the embryo quickly. Discharge yolk and other contaminants from the needle while keeping the PGCs.

Load clean silicone oil from the pool into the needle. Before collecting PGCs from a new embryo, deposit as much silicone oil as possible inside the somatic cell layer while keeping the loaded PGCs in the needle. After collecting PGCs from all embryos in a row, deposit the PGCs onto the surface of an embryo and unload any yoke or other contaminants onto another neighboring embryo.

In a similar manner, collect and combine PGCs from the embryos in the second row.