GeminiBio MXG102, Руководство пользователя

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Moxi GO II
™
User Guide
Page 58
5. When running future tests, verify the compensation setting is toggled to “Use Prior
Value” (other option is “None”). The system should automatically set this for you to
ensure that the established compensation value is applied to all future tests in the
experiment (until system power-off).
Note: If you power off the system, you will likely
need to reset the compensation value. In those cases, because the compensation
values have already been established, there is no need to run the single stain
controls. You can just run the next sample, touch compensation, and manually
adjust the values before saving (see below for more detailed instructions on
adjusting the compensation level)
Standard – Single Test - Compensation
On the Moxi GO II, data can be compensated by selecting the Comp icon at the bottom of a
new or saved test (image below – top/left ). Note: As would be expected, the compensation
icon is only available for tests in which multiple fluorescent channels have been recorded.
Selecting the compensation icon will load the quadrant-format scatter plot display of
fluorescence vs. fluorescence (image below top/middle and image below top/right).
Notes:
•
The only points displayed on this view are the points that were size-gated (between blue
markers) on the PMT vs size graph (image below – top/left).
•
In the vast majority of cases, spillover will occur from the green dye (PMT1) to the
orange/red channel (PMT2).
•
There is rarely to compensate for “backspill” from an orange/red dye (PMT2) into the
Green (PMT1) channel.
•
It may be necessary to select the Switch Axis button depending on the channel to be
compensated. The spillover channel (almost always PMT2) should always be set to the
y-axis. In the example below, PMT1 is originally on the y-axis (image below –
top/middle). To the switch axis button is selected to change/swap the axis (image below
- top/right) so that the FITC fluorophore (525/45nm PMT1, switched to x-axis) spillover
into the 561nm/LP channel (switched to y-axis) can be performed (image below –
top/right).
•
Initially, instructional text is displayed in red (over the scatter plot) and in yellow (on the
scatter plot). Once the gates are moved or compensation is adjusted, that text will
disappear until the user touches Clear (to remove changes) or Switch Axis.
When spillover occurs, there is a characteristic narrow slop to the data points on the graph
(images below – top/right and bottom/left). This occurs because of the direct correlation of a
fluorescent event on the green channel to a recorded spillover event on the red channel (i.e. a