Section 7
7
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Research Instruments Ltd
Hole Size Selection
For safe and effective treatment, we recommend that pulses used are in the range 15 to 1000µs.
Pulses that are very short may be ineffective and pulses that are too long may damage the embryo. For
this reason, a warning will appear on the
Fire
button when pulse width is 15µs or below and when the
pulse width is greater or equal to 1000µs.
When the warnings are shown, the laser can still be fired.
Ablation Procedure
Focus on the surface of an empty Petri dish and ensure that the target is correctly aligned to the pilot
laser spot. Place the dish with the specimen on the microscope stage and hold the specimen on the
bottom of the dish with a holding pipette. Check you are using the red Saturn 5 Laser System objective.
To ablate the zona, the focal plane must be at the mid section of the embryo. Refocus the microscope
until the image of the zona is sharp.
Focus here to ablate
Correct
Incorrect
X
Correct focus - specimen is sharply focussed.
Pilot laser brightness:
The brightness of the pilot laser can be adjusted by turning the control knob
on the rear panel (see Device Label
) to obtain the clearest image in the target alignment
procedure.
The light on the front of the control unit indicates that the control unit is operational. If the unit is
plugged in and the light is not on, contact Research Instruments for assistance.
Side view showing embryo on dish surface
