Burleigh PiezoDrill
Inertial Impact Drill User’s Manual
4 Recommended Procedure/Fluorinert
Burleigh Instruments, Inc. 2002 (08572-M-01 Rev D)
4-1
The information contained in this document is privileged information and
is intended for the personal and confidential use of Burleigh PiezoDrill customers.
4. Recommended Procedure for a Successful Injection with
Fluorinert in the Pipette
1.
Before starting the procedure, wash the end of the pipette in a solution of
medium with 10% polyvinylpyrolidone (PVP) added to it. The washing
process is accomplished simply by drawing a small amount of PVP
solution into the end of the pipette and then expelling it. This should be
repeated several times. The resultant coating on the end of the pipette
will help to prevent the pipette from getting sticky during the procedure.
It helps to wash the pipette periodically during the procedure as well.
Please review the following steps in conjunction with viewing the Nuclear
Transfer video on the enclosed CD.
2.
It is important to start with Fluorinert (FC-77) close to the tip of the
injection pipette when drilling is about to begin (see Figure 7). To do
this, apply positive pressure on the injection syringe. Adjust the pressure
until the Fluorinert is just inside the tip. The line inside the pipette that
moves in and out with the pressure is the meniscus.
Figure 7:
Initial setup for injection syringe
3.
Move the meniscus of Fluorinert close to the end of the pipette and
carefully pick up one sperm or nucleus in a microdrop with PVP medium
(with practice it is possible to load up the pipette with multiple sperm or
nuclei). Move to the microdrop without PVP, where the oocytes are, to
perform the transfer procedure. Apply positive pressure to reposition the
sperm or nucleus close to the tip of the pipette (note position in Figure 8
and the video).
Figure 8:
Pre-injection setup of injection pipette
