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Chapter 6 Troubleshooting
6
Troubleshooting chip images using the Chip View
Observation
56
Possible Cause
• The chip leaked during
thermal cycling or
imaging.
• A large bubble was
present in the chip
(insufficient Immersion
Fluid).
• Excess Immersion Fluid
is present on the chip
lid.
QuantStudio
Action
If you use the chip loader to seal the
chips, ensure that you are using at
least 20 lbs of pressure when applying
the chip lid. If you are not sure that
you are applying sufficient force,
increase the duration of the press.
Ensure that the ramp rate of the
thermal cycler is set to 1.6°C/second.
Ensure that the heated cover is set to
70°C (not to the default setting of
104°C).
If the steps above do not resolve
leaking, it may be beneficial to degas
the Immersion Fluid immediately
before use:
1. Ensure that the cap is on
securely.
2. Pull the syringe plunger to 3 mL
to create a partial vacuum in the
syringe.
3. Hold the syringe plunger at the
3-mL mark and shake vigorously
for ~30 seconds.
4. Hold the syringe vertically (tip
up), remove the cap, then use
the plunger to push any air out of
the syringe.
5. Use the Immersion Fluid
immediately.
If you are sealing chips manually, also
do the following to minimize leakage:
• Wear correctly fitted gloves to
prevent the material from
snagging during lid application.
• Ensure that the chip lid is
correctly aligned on the chip.
• Firmly press all four corners
when applying the lid.
• When pressing the chip, firmly
hold the sides of the chip and
wiggle it back and forth.
™
3D Digital PCR System User Guide
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