Absorbance Liquid Tests | 83
Synergy HT Operator's Manual
• Accuracy Specification. The following accuracy specifications are
applied using Normal mode and a 96-well microplate:
± 1.0% ± 0.010 OD from 0.000 to 2.000 OD
± 3.0% ± 0.010 OD from 2.000 OD to 3.000 OD
Liquid Test 2
The recommended method for testing the instrument’s alignment, repeatability, and
accuracy is to use the Absorbance Test Plate (see page 70). If the Test Plate is not
available, however, Liquid Test 2 can be used for these tests.
Required Materials
• A new 96-well, flat-bottom microplate, such as Corning Costar
®
#3590
• Ten test tubes, numbered consecutively, set up in a rack
• Calibrated hand pipette (Class A volumetric pipette recommended)
• Stock solution A or B (see page 81)
• A 0.05% solution of deionized water and Tween
®
20
Prepare the Dilutions
Refer to the table below and:
• Create a percentage dilution series, beginning with 100% of the original
concentrated stock solution (A or B) in the first tube, 90% of the original
solution in the second tube, 80% in the third tube, all the way to 10% in the
tenth tube.
• Dilute using the 0.05% solution of deionized water and Tween 20.
Table 2: Test Tube Dilutions for Liquid Test 2
Tube Number:
1 2 3 4 5 6 7 8 9 10
Volume of Original
Concentrated Solution (mL)
20
18
16
14
12
10
8 6 4
2
Volume of 0.05% Tween
Solution (mL)
0
2
4
6
8
10
12 14 16
18
Absorbance expected if original
solution is 2.0 at 200 µL
2.0
1.8
1.6
1.4
1.2
1.0
0.8 0.6 0.4
0.2
Note: The choice of dilutions and the absorbance of the original
solution can be varied. Use Table 2 as a model for calculating the
expected absorbances of a series of dilutions, given a different
absorbance of the original solution.
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