background image

Burleigh PiezoDrill

 Inertial Impact Drill User’s Manual 

2 Equipment and Materials

 Burleigh Instruments, Inc. 2002  (08572-M-01  Rev D) 

2-5

The information contained in this document is privileged information and

is intended for the personal and confidential use of Burleigh PiezoDrill customers.

Holding Pipettes

Manufacture or Purchase of Holding Pipettes

The Humagen price list in Appendix A includes holding pipettes with 85 

µ

m

and 120 

µ

m outside diameters.  The 85 

µ

m OD pipette is made for mouse

oocytes; the 120 

µ

m OD pipette is intended for swine and bovine oocytes.  It

is important that the outside diameter of the holding pipette be slightly
smaller than the oocyte.  This allows the oocyte to be held into position and
anchored to the slide or dish surface.  “Pinning” the oocyte in this manner
prevents the oocyte from moving when PiezoDrill vibrations begin.  The
inside diameter of the holding pipette should be large enough that some
cytoplasm is held in place by the holding pipette.  We recommend an ID of
20 to 25 

µ

m because:

It provides more perivitelline (PV) space between the zona and the oolemma
at the other side of the oocyte where the PiezoDrill will penetrate the zona.
Since zona vibration settings are higher than the settings for the oolemma,
the pipette must not engage the oolemma while vibrating at the zona settings.
PV space allows the zona to be penetrated with no possibility that the
oolemma will be penetrated accidentally at the same time.

It is easier to break the oolemma at low vibration settings when the
cytoplasm is held in this manner, which improves oocyte viability.

Figure 3 illustrates the correct size and position of the holding pipette.  Note:
This pipette has a slightly larger outer diameter than what we recommend.
Because this particular pipette is not bent and approaches the oocyte from
above, it still works effectively to pin the oocyte.

NOTE 

We have included one Humagen holding pipette with the PiezoDrill for
your initial use.

Summary of Contents for PiezoDrill

Page 1: ...User s Manual PiezoDrill Inertial Impact Drill 08572 M 01 Rev D Copyright Burleigh Instruments Inc 2002 All Rights Reserved PIEZODRILL is a trademark of Burleigh Instruments Inc...

Page 2: ......

Page 3: ...up 3 1 Filling the Injection Pipette with Fluorinert 3 1 Using Mercury in the Pipette 3 2 Attaching the PiezoDrill to the Microscope and the Micromanipulator 3 3 Critical Success Factors 3 4 PiezoDril...

Page 4: ......

Page 5: ...on the quality of the oocytes and many other factors that are beyond our control WARNING MERCURY WARNING USERS WHO CHOOSE TO USE MERCURY IN THE INJECTION PIPETTE WITH THE PIEZODRILL ARE STRONGLY CAUTI...

Page 6: ......

Page 7: ...rating the PiezoDrill and the following types of equipment Equipment type Equipment examples 1 Inverted microscope Leica DMIRB E Nikon TE200 300 Olympus IX70 Zeiss Axiovert 135 2 Injection pipette Hum...

Page 8: ...actor for productive PiezoDrill utilization If the pipette is not shaped correctly the procedure will not be successful PiezoDrill pipettes manufactured to Burleigh specifications can be purchased fro...

Page 9: ...optics at high magnification 300X 500X Any rough edges remaining after the pipette is broken should be gently removed by heat polishing using low heat The tip of the pipette does not need to be acid...

Page 10: ...indicates that heavier fluids in the pipette work better with the PiezoDrill Some researchers have used mercury for PiezoDrill procedures However due to the significant health risks associated with ex...

Page 11: ...e holding pipette should be large enough that some cytoplasm is held in place by the holding pipette We recommend an ID of 20 to 25 m because It provides more perivitelline PV space between the zona a...

Page 12: ...Holders Hollow tube systems see Figure 4 allow the hydraulic fluid used to control the movement of material to pass through the rod and enter directly into the pipette via an O ring type of holder at...

Page 13: ...the PiezoDrill and the syringe Solid Pipette Holders Solid pipette holders see Figure 5 require tubing from the syringe to be coupled directly to the pipette using Teflon or polyethylene tubing With...

Page 14: ...that the syringe responds precisely to fingertip commands If pressure inadvertently increases during the procedure Fluorinert will be pushed out of the end of the pipette If pressure inadvertently dec...

Page 15: ...s the Narishige B 8 ball joint see the PiezoDrill Operating Manual for assembly instructions The Narishige Adapter provides a stiffer connection that improves PiezoDrill performance We also make a Lei...

Page 16: ......

Page 17: ...the butt end of the pipette into the overhanging tubing 2 Firmly depress the syringe plunger allowing the Fluorinert to displace the air in the pipette thus filling the pipette Depending on the size...

Page 18: ...se or recommend the use of mercury in the PiezoDrill Mercury should not be used with the PiezoDrill by any individuals or laboratories unwilling to bear the entire risk of exposure to mercury or unabl...

Page 19: ...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...

Page 20: ...on the injection pipette side of the microscope check the following The pipette must be held tightly in the pipette holder The pipette holder must be tightly clamped in the PiezoDrill The pipette hold...

Page 21: ...5 Duration 60 66 Frequency 2 1 Bandwidth 50 50 It is evident from these settings that the use of mercury in the pipette causes much stronger pulses and requires much lower settings The bandwidth setti...

Page 22: ...ve one sharp pulse of the PiezoDrill with the following setting ADJUSTMENT SETTING Amplitude 35 Duration 90 Frequency 9 Bandwidth 100 Burleigh Instruments Inc does not suggest approve endorse or recom...

Page 23: ...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 c...

Page 24: ...nterline 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 pipett...

Page 25: ...r side of the embryo and activate pulse 2 to break the cytoplasmic membrane note in the video how the edge of the membrane next to the zona relaxes Using positive pressure inject the sperm or nucleus...

Page 26: ......

Page 27: ...use of Burleigh PiezoDrill customers 5 Recommended Procedure for a Successful Injection with Mercury in the Pipette The same 10 steps apply when using mercury in the pipette However only a small amoun...

Page 28: ......

Page 29: ...install the Narishige adapter in place of the ball joint 2 Be sure all manipulator and adapter connections are tight 3 Clamp the pipette holder on the manipulator as closely as possible to the PiezoD...

Page 30: ...cy settings on the PiezoDrill Oocyte lyses during zona drilling 1 Use less suction 2 Create more PV space see Holding Pipette section 3 Lower the amplitude and or frequency settings Some degree of lys...

Page 31: ...n Price List Burleigh Instruments Inc 2002 08572 M 01 Rev D A 1 The information contained in this document is privileged information and is intended for the personal and confidential use of Burleigh P...

Page 32: ......

Page 33: ...Burleigh Instruments Inc 7647 Main Street Fishers Victor New York 14564 8909 Tel 585 924 9355 Fax 585 924 9072 Email info burleigh com http www burleigh com PiezoDrill User s Manual 08572 M 01...

Reviews: