
Chapter 3 Scanning Operations
3-18
Applied Biosystems
3
Select the Gain
1.
Click the down arrow in the Gain field to display the list
of choices.
2.
Highlight the value you want (0–99). As previously
described in the “Gain” section, start off with a low value
and gradually increase the gain until you find the optimum
level(s) for your assay. Gain settings between 30 and 80
usually give the best performance. If you must use a value
below 30, your fluorophore is very concentrated and you
should dilute it. If you must use a value above 80 to detect
very low concentrations, try increasing the Reads/well
value or adjusting your assay design. (See “Reads/Well”
on page 3-9 for details.)
NOTE: When you create a file for a new assay, define
all four scans with the same filter pairs, but different
gain settings (from low to high). This way, you can
quickly determine the best gain setting for your
particular experiment.
Additional scans
Repeat the steps to select the number of scans, filter pair, and
gain for each additional scan you want to run.
Copying filter and
gain settings
You can copy the filter and Gain settings from a scan to up to 4
additional scans. For example, when you scan an entire 384
well plate, you must set up four scans per cycle. The
CytoFluor scanner reads 96 wells per scan. All four of the
scans must have the same filter settings and gain. You can set
the filter settings and gain for the first scan and then copy
those settings for the remaining scans using the CopyFilter To
option.
To use the Copy Filter To option:
1.
Set the filters and gain as described in “Select the filter
pairs for a Scan” on page 3-16 and “Select the Gain” on
page 3-18.
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