Running an Experiment
166
0112-0109 H
3.
Perform a
Protocol Signal Test
to evaluate the basal
fluorescence signal. The desirable fluorescence signal for
intracellular calcium assays is 200–1,200 fluorescence counts
for EMCCD camera and 500–5000 for ICCD camera.
When running a membrane potential assay, the basal
fluorescence counts should be between 1,500–2,500 counts for
standard, EMCCD camera or 10,000–15,000 for aequorin ICCD
camera to allow for a decrease or increase in the fluorescence
signal.
4.
If the basal fluorescence signal is substantially out of these
ranges, the excitation intensity, exposure time and gain or gate
can be adjusted. Use the suggestions below to adjust the basal
fluorescence.
Basal fluorescence too low:
•
Increase the
Excitation Intensity
if the basal fluorescence
signal is too low. During the assay, the
Excitation Intensity
should range between 20% and 100%.
•
Increase the Exposure Time. If you increase the
Exposure
Time
, the read interval will have to be increased to a minimum
of exposure time + 0.1 s (it takes 0.1 s to integrate the data).
•
Increase the
Camera Gain
(EMCCD camera). The gain can vary
between 1 (extremely bright fluorescence) to 240
(luminescence).
•
Increase the
Gate Open
(ICCD camera). The gate can vary
between 0.06% (extremely bright fluorescence) and 35% (very
dim fluorescence).
Camera Type
Calcium Assay
Membrane Potential
Assay
EMCCD
ICCD
EMCCD
ICCD
Excitation wavelength
(nm)
470–495
470–495
510–545
510–545
Emission Wavelength
(nm)
515–575
515–575
565–625
565–625
Excitation Intensity
(%)
50
50
50
50
Exposure Time (s)
0.4
0.533
0.4
0.533
Camera Gain
130
2000
50
2000
Gate Open
—
6%
—
6%
Содержание FLIPR Tetra
Страница 1: ...FLIPR Tetra High Throughput Cellular Screening System User Guide 0112 0109 H December 2011...
Страница 12: ...Contents 12 0112 0109 H...
Страница 16: ...System Overview 16 0112 0109 H...
Страница 40: ...System Hardware Features 40 0112 0109 H...
Страница 148: ...Exchanging Hardware 148 0112 0109 H...
Страница 156: ...Calibration and Signal Test 156 0112 0109 H...
Страница 196: ...Running an Experiment 196 0112 0109 H...
Страница 232: ...Robotic Integration 232 0112 0109 H The following drawings illustrate these requirements...
Страница 282: ...Data Processing Algorithms 282 0112 0109 H...
Страница 294: ...Consumables and Accessories 294 0112 0109 H...
Страница 298: ...Using AquaMax Sterilant 298 0112 0109 H...
Страница 302: ...Electromagnetic Compatibility EMC 302 0112 0109 H...