Bio-Rad Bio-Plex® Assay Builder - Manual (Page 12)

6. If acquiring data from more than one plate, empty the waste bottle
and refill the sheath bottle after each plate. Select Wash Between
Plates and follow the instructions for fluidics maintenance. Then
repeat the Prepare Protocol and Acquire Data steps.
7. When data acquisition is complete, select Shut Down and
follow the instructions.
Reacquire Data
It is possible to acquire data from a well or plate a second time using the
Rerun/Recovery mode located below Start in Step 7 (Run Protocol).
1. Check the wells where data will be acquired a second time.
Any previous data will be overwritten.
2. Remove the buffer by vacuum filtration and add 125 µl of
resuspension buffer to each well. Cover the filter plate with a new
sheet of sealing tape.
3. Repeat Acquire Data steps 1–6 to acquire data a second time.
The data acquired should be similar to the data acquired initially;
however, the data acquisition time will be extended since fewer
beads are present in each well.
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Acquire Data
1. Shake the assay plate at 1,100 rpm for 30 sec immediately before
acquiring data. Failure to do so will result in increased data
acquisition time due to bead settling.
2. Check that the filter plate is flat. While pressing on one end of the
plate, observe the distance that the opposite end of the plate is
raised off a flat surface. If the distance is >1 mm, transfer all contents
to a flat-bottom 96-well plate or another filter plate.
3. Visually inspect the plate and ensure that the assay wells are filled with
buffer prior to placing the plate in the Bio-Plex microplate platform.
4. Slowly remove the sealing tape and any plate cover before placing
the plate in the reader.
5. Select Step 7 (Run Protocol):
a) Specify data acquisition for 25 beads per region.
b) In Advanced Settings, confirm that the default DD gate values
are set to 4335 (low) and 10000 (high).
NOTE: When using a Luminex 100 instrument, set the gates
according to the Luminex procedure located in the manual.
c) Select Start and save the .rbx file. Then follow the instructions for
data acquisition.
Sample Information Example