Burleigh PiezoDrill
Inertial Impact Drill User’s Manual
4 Recommended Procedure/Fluorinert
4-2
(08572-M-01 Rev D)
Burleigh Instruments, Inc. 2002
The information contained in this document is privileged information and
is intended for the personal and confidential use of Burleigh PiezoDrill customers.
4.
With the embryo on the holding pipette pinned to the dish, approach the
injection pipette to the embryo as shown in Figure 9, deflecting it
slightly.
5.
When the injection pipette touches the zona, apply negative pressure in
the injection pipette. The sperm or nucleus will move away from the
oocyte, creating a seal between the injection pipette and the zona. The
seal is where the drilling occurs (as shown in Figure 9 and the video).
NOTE
It is very important to be at the centerline vertically
and
horizontally. If
the embryo jumps around while drilling, the injection pipette likely is
not centered in the vertical plane.
Figure 9:
Pre-drilling position of the injection pipette
6.
Activate pulse 1 on the drill;
do not push the injection manipulator at
this point – let the PiezoDrill do the work
. Increase the negative pressure
slightly as the tool drills through the zona (as shown in Figure 10), being
careful not to break through the oolemma. (In the video, note the
movement of the nucleus away from the oocyte as drilling occurs.) Near
the point of final penetration, decrease the suction slightly. After cutting
through the zona, stop the drill and move the pipette to the PV space to
ensure that it is in.
Figure 10:
Initial drilling
7.
Remove the tool and apply positive pressure on the syringe until the core
(removed material) comes out the tip as shown in Figure 11. The
cohesiveness of the core will vary slightly depending on species and
other factors.
