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9 changes: 4 additions & 5 deletions 03_flow_MEFL/exercises.m
Original file line number Diff line number Diff line change
Expand Up @@ -71,8 +71,7 @@
CM = ColorModel(beadfile, blankfile, channels, colorfiles, colorpairfiles);
CM = set_ERF_channel_name(CM, 'FITC-A'); % Name the channel we'll use for ERF units
CM=set_dequantization(CM, 1); % important at low levels
CM=set_bead_plot(CM, 2); % 2 = show beads for all channels, even though only FITC will be used
CM=set_bead_min(CM, 1); % Don't consider beads less than this amount
TASBEConfig.set('beads.rangeMin', 1); % Don't consider beads less than this (log10) amount
% Things we'll talk about in the next section...
CM=set_translation_plot(CM, true);
CM=set_translation_channel_min(CM,[2,2,2]);
Expand All @@ -88,15 +87,15 @@
% The FITC channel is the one that we will key off
% The identification is pretty terrible, though!
% Two things are going wrong here:
% 1) We're getting smearing of peaks from autofluorescence: bead_min should be raised
% 1) We're getting smearing of peaks from autofluorescence: beads.rangeMin should be raised
% 2) Automatic threshold detection is not finding the right value, because there
% is not a distinct enough "valley" in the FITC graph
% Notice also that you got a warning that: "Warning: Bead calibration probably incorrect"
% When peaks are mis-detected, this typically leads to a bad fit against the
% expected sequence gaps, giving warning of failures.

CM=set_bead_min(CM, 2); % Don't consider beads less than this amount
CM=set_bead_peak_threshold(CM, 200); % override default peak threshold
TASBEConfig.set('beads.rangeMin', 2); % Don't consider beads less than this (log10) amount
TASBEConfig.set('beads.peakThreshold', 200); % override default peak threshold
CM = resolve(CM);

% We get a new warning: "Warning: Only one bead peak found, assuming brightest"
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