Fluorescence Tests | 117
Important! The tests presented in this section require
specific microplates, solutions, and EX/EM filters. Your
laboratory may require a deviation from some of these tests.
For example, you may wish to use a different fluorescing
solution and/or microplate.
If deviation from the tests as presented in this section is
required, the following steps should be taken the first time
each test is run (e.g., during the Initial OQ):
1. Perform the tests exactly as described on the following
pages.
2. Rerun the tests using your particular solutions, filters,
microplates, etc. If results are comparable, then the results
from these tests will be your baseline for future tests.
3. Be sure to document your new test procedure(s), and save
all test results.
Required Materials
BioTek offers a liquid test kit (PN 7160013) containing the microplates
and solutions used in this procedure.
Microplates should be perfectly clean and free of dust or bottom
scratches. Use new microplates from sealed packages.
Manufacturer part numbers are subject to change.
Methylumbelliferone can be used as an alternative or supplemental
method for performing these tests for the top probe. See the
instructions starting on page 123.
• Buffer:
NIST-traceable Sodium Borate Reference Standard (pH 9.18) (e.g., Fisher-
Scientific 1 L Sodium Borate Mfr. #159532, or equivalent),
or
Phosphate-Buffered Saline (PBS), pH 7.2-7.6 (e.g., Sigma tablets, Mfr. #P4417,
or equivalent) and pH meter or pH indicator strips with pH range 4 to 10
• Sodium Fluorescein Powder (1 mg vial, BioTek PN 98155)
• Bottom optics: A clean Hellma Quartz 96-well titration plate (Mfr.
#730.009.QG), or equivalent, such as the 96-well glass-bottom Greiner
SensoPlate (Mfr. #655892).
• Top optics: A new, clean, 96-well solid black microplate, such as Corning
Costar Mfr. #3915. The Hellma Quartz plate described above can also be used.
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