Biotek Synergy LX, User Manual

Page: 91 / 198

67 | Chapter 5 : Instrument Qualification Process
Absorbance Liquid Tests
BioTek Instruments, Inc. has developed a series of liquid test procedures for testing your
reader's absorbance system.
Test Methods
Absorbance Liquid Test 1 confirms repeatability and alignment when a solution is used in
the microplate. If these tests pass, then the lens placement and optical system cleanliness
are proven. For the Repeatability portion of this test, two columns containing a color-
absorbing solution are read five times at 405 nm. For each well, an "allowed deviation" is
determined based on its Mean OD and the reader's repeatability specification. Each well's
Standard Deviation must be less than its Allowed Deviation to pass. To confirm the reader’s
mechanical alignment, the plate is rotated 180 degrees in the carrier (e.g., A1 is now in the
H12 position), and the same two columns are read. The initial and new OD readings are
compared using the reader's accuracy specification. If the two readings in the same well do
not meet specification, the reader may be out of alignment.
If an Absorbance Test Plate is not available, Absorbance Liquid Test 2 may be conducted to
test the instrument's alignment, repeatability, and accuracy by preparing a series of
solutions of varying OD values as described on page 94.
Absorbance Liquid Test 3 is an optional test offered for those sites that must have proof of
linearity at 340 nm. (Alternatively, the BioTek 340 nm Absorbance Test Plate may be used;
see page 64.) This test is optional since the
Synergy LX has good “front end” linearity
throughout the specified wavelength range. While the absolute values of the OD cannot be
determined by this test, the results will indicate if there is adequate repeatable absorbance
and a linear slope. This method is dependent upon proper dye dilution and a skilled
pipetting technique. It is expected that the first dilution (mid-level solution) will have an
absorbance value near 75% of that of the stock (high-level) solution and that the second
dilution (low-level solution) will have an absorbance value near 50% of that of the stock
solution.
Gen5 Protocol Parameters
The information in this section represents the recommended reading parameters for the
referenced Gen5 protocol(s). It is possible that your tests will require modifications to
some of these parameters, such as the Plate Type.
The Plate Type setting in each Gen5 protocol should match the actual 96-well plate in
use.
Synergy LX User Manual
1501000N Revision A BioTek Instruments, Inc.