Molecular Devices FLIPR Tetra, User Manual

Page: 161 / 302

FLIPR
®
Tetra High Throughput Cellular Screening System User Guide
0112-0109 H 161
8. Set the Camera Gain to 80 (EMCCD camera).
9. Set the Gate Factor to 3% (ICCD camera).
10. Place the yellow signal test plate on the stage in the read
position.
11. Take a picture by clicking Yellow Plate Signal Test . Acceptable
test plate results are a standard deviation less than 5% when
the instrument is calibrated with the yellow test plate for the
appropriate plate format. If the test results are outside of the
range of acceptable values, look at the Image Display to see if
any anomalies can be found on the plate. For additional
information see, Chapter 9: Troubleshooting on page 197 .
12. Print the results and keep them in a “Maintenance” folder by the
instrument to track the standard deviation of the test plate over
time. Alternatively, you can save the files on the hard drive in
the signal test directory ( C:\Documents and
Settings\[your_user_name]\My Documents\Molecular
Devices\ScreenWorks\MySignalTests ) as a * .sig file.
Dye-Loading the Cells for Fluorescence Assays
Many cell-based responses require fluorescent dye loaded in cells to
bind or change conformation in the presence of a ligand. These changes
are monitored through a shift in emission wavelengths and captured by
a CCD camera. Calcium sensitive dyes such as the ones used in Molecular
Devices
FLIPR
®
Calcium Assay kits monitor intracellular calcium flux assays,
or environmentally sensitive for membrane potential assays. Depending
on the application evaluated, dye-loading typically lasts 30 min–1 hour
(at either room temperature or 37 °C).
Loading Duration and Temperature
Optimal loading time depends on the cell type and presence of an anion
exchange inhibitor. It is recommended not to exceed the optimal
loading time. If anion exchange inhibition does not enhance
dye-loading conditions, it should not be used. A 60 minute loading time
Note: The Yellow Plate Signal Test and Image Display results are
not saved within a data file.
You do not have to run a yellow plate signal test more than once a day,
unless you recalibrate or change the optics.
This Fluorescence signal test also checks the Luminescence optics,
since the optics path is the same in both modes.