Qiagen QIAcuity 911000 User Manual page 210

Reference channel
The Software Suite also provides a 1D Scatterplot view for the reference channel, which is hidden
by default. To view the 1D Scatterplot for the reference channel in addition, click Show reference
channel. The title of this 1D Scatterplot view indicates that the 1D Scatterplot is related to the
reference channel. The graph for the reference channel provides the possibility to set a lower as
well as an upper threshold to exclude partitions with too high/too low RFU. Setting the thresholds
(individually per well or common lower and upper threshold for all selected wells) is analog to 1D
Scatterplots for target channels. To apply the new threshold values for the analysis, click Recalculate.
The 1D Scatterplot for the reference channel uses the same functions as the 1D Scatterplot views for
non-reference channels.
1D Scatterplot for the reference channel.
Lasso mode for partition assignment
Alternative of changing threshold values the user can assign partitions using a free hand selection
in the 2D Scatterplot view (Lasso mode). When the user marked points by lasso for two channels
or targets all thresholds on 1D Scatterplot for them disappear and can't be modified before
switching back to threshold mode using the Auto-threshold button. When the user clicks recalculate
button, changes in threshold selection affects 2D Scatterplot results as well.
Channels which are not used in 2D Scatterplot with lasso mode do still show the threshold mode
in the 1D Scatterplot view.
Refer to 2D Scatterplot tab for absolute quantification for more details regarding the assignment of
partitions using the lasso mode.
QIAcuity User Manual 06/2022 210