Molecular Devices FLIPR Tetra, User Manual

Page: 175 / 302

FLIPR
®
Tetra High Throughput Cellular Screening System User Guide
0112-0109 H 175
monolayer are recommended to ensure better mixing of
compounds and lower signal variance across the plate. However,
further assay development, adjustment of the volume, height
and speed of dispense, is recommended to optimize your cell
response.
Recommended Setup Parameters
Recommended experimental setup parameters are as follows.
Experimental Setup
Parameters
EMCCD
Cameras
ICCD Camera
Read Mode Fluorescence Fluorescence
Excitation Wavelength 470–495 nm 470–495 nm
Emission Wavelength 515–575 nm 515–575 nm
Excitation Intensity 80%
1
50
1
%
1. Can be adjusted to increase or decrease the signal if low RFUs or saturation
problems occur.
Camera Gain 50–130 2000
Gate Open — 6
1
%
Exposure Length 0.4
1
s 0.53
1
s
Read Interval 1 s 1 s
Addition
Parameters
96-Well
Plate
384-Well
Plate
1536-Well
Plate
Compound
concentration (fold)
5X 5X 7X
Addition volume ( μ L) 50 12.5 1
Addition height ( μ L) 210–230 25–45 1–2
Tip up speed (mm/s) 10 10 5
Dispense speed ( μ L/s)
Adherent cells
50–100 30–40 4–7
Dispense speed ( μ L/s)
Non-adherent cells
10–20 10–20 1–5