Burleigh PiezoDrill
Inertial Impact Drill User’s Manual
3 Setup
Burleigh Instruments, Inc. 2002 (08572-M-01 Rev D)
3-3
The information contained in this document is privileged information and
is intended for the personal and confidential use of Burleigh PiezoDrill customers.
through the back-end of the pulled pipette, leaving about ¼ inch long strip
inside the pipette. Mount the pipette onto the injector. Push mercury down
to the tip. There maybe a small air bubble between the oil (or water) and the
mercury. Draw some media through the tip of the pipette so that the media
provides a protective barrier between the mercury and the tip of the pipette.
You are now ready to draw sperm cells into the pipette to begin the ICSI
process.
After a few sperm injections, the cytoplasm from the egg begins to build-up
on and inside the tip of the injection pipette. The pipette becomes sticky and
it becomes impossible to move the sperm heads in and out smoothly.
Therefore, it is necessary to frequently "wash" the pipette tip. It is good
practice to have a drop of medium in the ICSI dish that is used strictly as a
wash drop. Move the pipette into this drop and eject a little bit of mercury
from the tip. Actually, it is washed in and out but every time the pipette is
drawn back up, whatever mercury was ejected will break off and form a
bubble in the media drop. Once outside the pipette, the mercury will not go
back through the fine tip of the pulled pipette, resulting in a medium drop
with many little bubbles of mercury.
For this reason, all of the ICSI dishes must be considered mercury
contaminated. All oil, pipettes and all ICSI dishes must be disposed of as
hazardous waste and labeled as containing mercury. Place used pipettes into
a 50-ml plastic tube for containment and disposal. Even the water or oil in
the tubing that attaches to the microinjection system must be considered
mercury contaminated. Drips on the counter can't be placed into regular
trashcans; they are mercury contaminated and should go into hazardous
waste receptacles.
Keep a vacuum flask near where the mercury is used and stored. If a spill
occurs, use a regular household-type hand vacuum with a plastic tube
attached to the vacuum bottle to clean all potentially contaminated surfaces.
Attaching the PiezoDrill to the Microscope and the Micromanipulator
Place the PiezoDrill control unit so that the controls can be easily reached
while sitting at the microscope. The PiezoDrill is equipped with foot pedal
controls that allow two independent sets of parameters to be readily accessed
via independent foot pedals. This feature is important since different
parameters will be employed when drilling the zona pellucida versus
breaking the cytoplasmic membrane (oolemma). Insert the pipette holder
into the PiezoDrill according to the instructions in the Operating Manual,
then clamp the pipette holder onto the manipulator. It is desirable to clamp
the pipette holder as closely as possible to the PiezoDrill (see examples in
Figures 4 and 5). Also, it is important that all connections, including the
Narishige or Leitz adapter, if used, are completely tightened. This type of
attachment will ensure optimum vibrations at the pipette tip.
