For an overview of biexponential transformation, click here. Click the transform (T) button on the graph window.įor more details, including changing Transformation values, click here. ![]() To adjust the Transformation for any parameter, select a parameter in the graph’s axis label drop down list. The sample was blank beads plus a few contaminants.Ī biexponential transform will be applied to all digital data (fcs3.0 data files) based on the cytometer that the data was acquired on. The data below was collected with the FACSDiva software. Only the scales have been altered to display the double negative population as a cluster. If 1,000 cells are represented as double negative by a plot that has picket fencing, 1,000 cells are represented by a plot that has been visually smoothed by applying the biexponential transformation.īelow is a figure showing a normal plot on the left and a transformed plot on the right. It is important to note that transforming the data does not alter data in any way. broken lines displayed on digital data around zero) can be visually smoothed using FlowJo’s display biexponential transformation feature. For a description of the differences between analog and digital data, see the section “Analog and Digital Data” below. This discrepancy gives rise to visual artifacts in graphs such as picket fencing. Events with low signal intensity (near zero) generally have lower resolution than events appearing higher on the scale when viewed on a logarithmic scale. The mathematics behind this conversion is complicated but can be reduced to the amount of resolution a cell or “event” has when its information is recorded. ![]() Transform digital data quickly and easily in FlowJo using the transform function on the graph window.įlow cytometers collect analog information and convert it to digital information via hardware and software components during acquisition.
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