Bio-Rad Bio-Plex® 200 Systems - Manual (Page 30)

1 Make sure that the system has been set up according to the directions in the Millipore user
guide.
Hint: Make sure that the connector leading to the vacuum control knob is perpendicular to
the manifold. This will ensure that no buffer travels to the vacuum control knob.
2 Place a 96-well flat-bottom microplate (not a filter plate) on the vacuum apparatus.
3. Make sure the gold vacuum control valve (Figure 23) is completely open (all colors
showing).
4. Make sure the gray ON/OFF valve (Figure 24) is completely off (the knob should be
perpendicular to the direction of the tubing attached to it).
Fig. 23. Gold vacuum control valve. Fig. 24. Gray ON/OFF valve.
5. Turn on the lab vacuum to maximum level and note this setting for performance of assays.
In common lab vacuum systems, the lever should be turned so that it is parallel to the
vacuum port. Alternatively, turn on the vacuum pump.
6. Slowly open the ON/OFF valve on the manifold to the ON position (parallel to the tubing).
7. Slowly turn the vacuum control valve clockwise until a slight vacuum sound is heard.
8. Press firmly on the 96-well plate (all four corners) until it is sealed securely by the vacuum
on the apparatus.
9. Observe the pressure reading on the attached gauge (see below).
10. Adjust the ON/OFF valve and turn the calibration knob so that the gauge reads approximately
1–2.5" Hg.
11. Close the lab vacuum until the 96-well plate is released from the apparatus.
12. Turn on the lab vacuum again, press on the 96-well plate, and look at the indicated pressure
on the gauge again.
13. If the pressure is still approximately 1–2.5" Hg, the vacuum apparatus has been correctly
calibrated. If not, repeat the steps of the calibration procedure until the desired result is
achieved.
14. Turn off the vacuum apparatus and proceed to Section 3.12.2, Validation of Vacuum
Pressure.
Note: Do not make any further adjustments to the vacuum control valve.
3.12.2 Validation of Vacuum Pressure
Validating the Vacuum Manifold Pressure for a Bio-Plex Assay
1. Prewet all of the wells of a Millipore 96-well filter plate with 100 µl PBS.
2. Place the prewetted filter plate on the calibrated vacuum apparatus.
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