Bio-Rad iQ™5 Optical System Software, Version 2.1 - Manual (Page 128)

Section 7 Calibrating the Instrument
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Select/Add Fluorophores
To choose the fluorophores needed to calibrate the instrument, select from the predefined
fluorophores or add custom fluorophores by clicking Select/Add Fluorophores. The Fluor Selector
will be displayed (Figure 7.8).
Fig. 7.8 The Select/Add Fluorophores Dialog Box
Add a fluorophore from the list displayed by clicking the checkbox next to the fluorophore name.
To add a custom fluorophore that is not on the list, select the appropriate filter pair from the
drop-down menu. Refer to section 9.2.1 for information about system filter specifications and
recommended fluorophores.
Specify the name and color of the custom fluorophore by typing the name in the Fluor Name field
and clicking Add New Fluor. The new fluorophore name appears in the list for the selected filter
pair. Click the checkbox next to the new fluorophore name, then select the color to specify the
fluorophore in the graphic by clicking on the color field that appears. Choose contrasting colors for
maximum clarity, then click OK.
NOTE: Two or more different dyes may be used with the same filter pair; however, two dyes
using the same filter pair may not be used in the same well.
Select new Sample Volume, Seal Type and Vessel Type from the drop-down menus as appropriate
for this experiment, then click Save & Exit Plate Editing. Choose a meaningful file name, and save
the Pure Dye Plate Setup.
IMPORTANT: Pure dye calibrations are made for each instrument, dye type, sample volume,
seal type, and vessel type, and are instrument- and experiment-specific. Do not attempt to use
pure dye calibration results on a different instrument, seal or vessel type, or sample volume.
7.5 Viewing Calibration Files
Background and well factors calibration files are saved to their respective folders within the iQ5
program folder. Pure dye calibration files are saved in the RMEData folder in the iQ5 program
folder. Previous calibrations are saved in the subdirectory called “Backup”. It is advisable to
periodically backup your calibration files by copying the calibration folders and their contents.