Luminex xPONENT 4.2 for FLEXMAP - Manual (Page 32)

Biological Samples
CAUTION: Human and animal samples may contain biohazardous
infectious agents. Where exposure to potentially biohazardous
material—including aerosol—exists, follow appropriate biosafety
procedures and use personal protective equipment such as
gloves, gowns, laboratory coats, face shields, or mask and eye
protection. Use ventilation devices. Observe all local, state, and
federal biohazard handling regulations when disposing of
biohazardous waste material.
Dilute concentrated biological samples, such as plasma or serum, at least 1:5 with reagents
as part of assay setup or as a final dilution step to reduce the chance of system clogs. If you
are running a MagPlex
® or MicroPlex ® kit, follow the dilution instructions in the kit’s product
insert.
Bead (Microsphere) Handling
MicroPlex ® and MagPlex ® beads come in various configurations. To reduce foaming and
precipitation, avoid agitating the beads until you are ready to vortex and use them. Beads
settle and must be resuspended by vortexing before use. In addition:
• Multiple pipetting from the original container can affect bead concentrations.
• Protect MagPlex ® and MicroPlex ® beads from light at all times to prevent photobleaching.
Photobleaching effects are cumulative. To maintain the integrity of the beads, minimize
their exposure to light.
• Store MagPlex ® and MicroPlex ® beads at 2 degrees C to 8 degrees C.
NOTE: Refer to the product information sheet that accompanies your
MapgPlex ® beads or assay for additional information.
Sample Volume
Sample volume or sample sizes range from 10 to 200 µL. Ensure that some of the sample
remains in the well after aspiration; about 25 µL greater than the sample volume. This
amount may vary depending on the type of plate used. After acquisition, the Luminex
®
analyzer washes the sample lines resulting in ejection of approximately 45 µL of sheath fluid
back into the well for a 96 and 384-well plate. Ensure that there is room to add this amount to
the well without overflowing and contaminating other wells.
The volume restrictions on the assay design can be expounded by the following formula:
Total well volume (µL) - Sample uptake volume (µL) + 45 (µL) <Maximum Well Volume (µL)
• Total well volume = Starting sample volume of a well before the unit samples for
acquisition. Well volume is determined by the consistency of the bead set.
• Sample uptake volume = Uptake volume for acquisition (program this in the protocol as
sample volume).
• 45 (μL) = Volume expelled back into the well.
xPONENT ® for FLEXMAP 3D ®
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