Molecular Devices FLIPR Tetra, Руководство пользователя

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Running an Experiment
174 0112-0109 H
Loading the Cells Using the Loading Buffer
1. Remove cell plates from the incubator or centrifuge. Do not
remove the supernatant. Add an equal volume of Loading Buffer
to each well (100 μ L per well for 96-well plates, 25 μ L for 384-
well plate).
2. Incubate cell plates for 1 hour at 37 °C and then keep the plates
at room temperature until used (loading time should be
optimized for your cell line).
CAUTION! Do NOT wash the cells after dye loading.
Running the FLIPR Calcium Assay Kit
To start the calcium assay:
1. After incubation, acclimate the plates to room temperature for
approximately 10–20 minutes prior to transferring them to the
FLIPR
®
Tetra System. Please refer to “Recommended Setup
Parameters” on page 136 for running the Calcium assay kit on
FLIPR
®
Tetra System.
2. When performing a signal test prior to an experiment, typical
average baseline counts range from 800–1,200 RFU for the
EMCCD camera and 500–5000 for the ICCD camera on FLIPR
®
Tetra System.
3. Suggested experimental setup parameters for each FLIPR
®
System are as follows. Fast addition speeds close to the cell
Note: Add 2 μ L per well for 1536-well plate by using an
AquaMax
®
DW4 or equivalent device.
Note: Although Molecular Devices does not recommend
washing cells before dye loading growth medium and serum
may interfere with certain assays. In this case, the supernatant
can be aspirated and replaced with an equal volume of serum-
free HBSS plus 20 mM HEPES buffer before adding the Loading
Buffer. Alternatively, cells can be grown in serum-free
conditions.
Note: Some assays perform optimally when the plates are
incubated at room temperature.