Biotek Synergy HTX, Operator'S Manual, Page: 131 / 184

Absorbance Liquid Tests |
109
Emission: Hole
Optics Position: Top
Gain/Sensitivity: 50
Read Height: 1.00 mm
Troubleshooting
If either test fails, try the following suggestions. If the test(s) continues to fail, print the
results and contact BioTek’s Technical Assistance Center.
• Ensure that the reading is performed through a hole in the filter wheel, not
through a glass filter.
•
Verify that the filter settings in Gen5 match the physical wheel.
• If the test continues to fail, the optical probe(s)
may
need to be cleaned.
Contact BioTek’s Technical Assistance Center for instructions.
Absorbance Liquid Tests
Conducting Liquid Tests confirms the Synergy HTX’s ability to perform to specification
with liquid samples. Liquid testing differs from testing with the Absorbance Test Plate in
that liquid in the wells has a meniscus, whereas the Test Plate’s neutral density glass filters
do not. The optics characteristics may differ in these two cases, thus alerting the operator
to different types of problems.
•
Liquid Test 1
confirms repeatability and alignment of the reader when a solution
is used in the microplate. If these tests pass, then the lens placement and optical
system cleanliness are proven.
•
Liquid Test 2
can be used to test the alignment, repeatability, and linearity of the
reader if an Absorbance Test Plate is not available.
•
Liquid Test 3
is provided for sites requiring proof of linearity at wavelengths
lower than those attainable with the Absorbance Test Plate. This test is optional
because the reader has good “front end” linearity throughout its wavelength range.
For Liquid Tests 1 and 2, instructions are given to prepare the stock dye solution described
on the next page. The purpose of the formulation is to create a solution that absorbs light
at ~2.0 OD full strength when dispensed at 200
µ
L in a flat-bottom microplate well.
Alternatively, any solution that gives a stable color will suffice. (This includes substrates
incubated with an enzyme preparation and then stopped with an acidic or basic solution.)
Some enzyme/substrate combinations that may be used as alternates to the described dye
are shown below:
Synergy HTX Operator's Manual