Biotek Synergy LX, User Manual

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Recommendations for Optimum Performance | 51
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When using absorbance, nonuniformity in the optical density of the well bottoms
can cause loss of accuracy, especially with U- and V-bottom polyvinyl microplates.
Check for this by reading an empty microplate. Dual wavelength readings can
eliminate this problem or bring the variation in density readings to within
acceptable limits for most measurements.
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Inaccuracy in pipetting has a large effect on measurements, especially if smaller
volumes of liquid are used. For best results in most cases, use at least 100 µL per
well in a 96-well plate.
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Pipetting solution into 384-well plates often traps air bubbles in the wells, which
may result in inaccurate readings. A dual-wavelength reading method usually
eliminates these inaccuracies. For best results, however, remove the air bubbles
by degassing the plate in a vacuum chamber or spinning the plate in a centrifuge
before reading.
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The inclination of the meniscus can cause loss of accuracy in some solutions,
especially with small volumes. Shake the microplate before reading to help bring it
within acceptable limits. Use Tween 20, if possible (or some other wetting agent)
to normalize the meniscus for absorbance measurements. Some solutions develop
menisci over a period of several minutes. This effect varies with the brand of
microplate and the solution composition. As the center of the meniscus drops and
shortens the light path, the density readings change. The meniscus shape will
stabilize over time.
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It is the user’s responsibility to understand the volumetric limits of the plate type
in use as it applies to the assay being run.
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Use of liquids with concentrations of acids, corrosives, or solvents of 3% and
greater can begin attacking the materials inside the instrument's chamber.
Running multiple plates with concentrations < 3% in long kinetic experiments may
also have a destructive effect. If the experiment is incubated, it will accelerate the
deterioration of chamber components. When in doubt about the use of acids,
corrosives, or solvents, please contact
Technical Support .
Using 384-Well Microplates
When using a 384-well microplate, you can use the Gen5 Auto Map feature to ensure you
are using an accurate plate map for your reads. See the Gen5 Help for more information.
Synergy LX User Manual
1501000N Revision A BioTek Instruments, Inc.