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Scales and transforms

Every axis has a scale, chosen independently of the parameter. Getting it wrong is the most common reason a plot looks unusable, and it is almost always mistaken for a gating problem.

Click an axis label to change the parameter and its scale.

Scale Use it for
Linear Scatter channels (FSC, SSC) and time. Almost never right for fluorescence.
Log Fluorescence with no compensation and no negative values. Cannot represent zero or negatives at all.
Logicle The usual right answer for compensated fluorescence — log-like at high values, linear near zero, so negatives from compensation still display.
Asinh Similar behaviour to logicle with a simpler parameterisation; a good default when logicle needs fiddling.
Hyperlog Another log-linear hybrid. Useful when logicle piles events against the axis near zero.

Compensation subtracts spillover, and subtraction produces negative values for dim and negative populations. That is expected, not an error.

A log scale cannot show a negative number. Events either vanish or jam against the axis, and a real negative population turns into a smear that looks like a staining failure. Logicle, asinh and hyperlog exist for exactly this: log-like where the data is bright, linear through zero where the data is dim.

If a population is piled against an axis or crushed into a corner, change the scale before you touch the gate.

The app suggests a scale per channel from the data itself, rather than defaulting everything to linear. Treat the suggestion as a good starting point: it is derived from the distribution, not from knowledge of your panel, so a channel with unusual staining may want overriding.

  • Scatter: linear.
  • Time: linear.
  • Compensated fluorescence: logicle, or asinh if logicle needs tuning.
  • Uncompensated single-colour, no negatives: log is fine.
  • Anything that looks wrong at zero: try hyperlog.

A gate is a region in whatever coordinate space the plot is showing. Change the scale or the compensation underneath it and the events falling inside can change.

Order that avoids surprises:

  1. Fix channel names and compensation.
  2. Choose scales.
  3. Gate.

If you must change a scale after gating, look at the affected gates rather than assuming they travelled with it.