Making Movies in FlowJo

You can generate a movie form from both the Graph Window and the Layout Editor. Movies made in the Graph Window display the data from a single population. Movies made in the Layout Editor display data from multiple samples.

Movies will be saved as Quicktime files. If you need Quicktime, please visit Apple's website here.

This feature provides an entirely new way to explore data.

Movies generated from the Graph Window

To generate a movie from a graph window select Make Movie from the Graph menu. You will be presented with the Movie Specification window. FlowJo will divide the events into slices that are ordered sequentially across the control parameter. The primary concern is to choose the control parameter, which is the third dimension that will be added to the graph. If time is an existing parameter in your data, then it will be the default choice as the control parameter. Otherwise, you can choose any other parameter you wish.

You can choose whether the slices will be made with an even number of channels per slice, (check Equal Number of Channels per Frame) or with a variable width slice that contains an equal number of events in each of the slices. (Remove the checkmark from Equal Number of Channels per Frame)

In the generated movie, the control parameter will be drawn in a histogram, with a red bar over the area that contains the events currently visible in the main (upper) graph. If the Equal number of channels per frame box is selected, the red bar will always have the same width as it moves across the histogram. If it not checked, the width of the red rectangle will vary so that the area under the curve inside the red bar is the same for each frame.

You can specify the Maximum number of frames, that the movie will contain. The larger the number of frames, the longer it will take to generate the movie, but the more sensitive it will be to trends that exist within a small range of values.

For some types of analysis, it is useful to graph events not of each individual slice, but for all of the events up to and including the current slice. The checkbox Display accumulates as movie progresses will cause the graph of each frame to include all of the events with a value for the control parameter of less than or equal to the frame's current value.

To get an idea of how these controls effect a movies, see these example movies:

• Time as the movie control: Kinetic display of calcium flux
• A third parameter as a movie control (equal channels per frame)
• A third parameter as a movie control (equal events per frame)

Movies Generated from Layout Editor

The movies generated in the Layout Editor are oriented towards visualizing trends or differences among samples as opposed to the Graph Window where the focus is on a single parameter in one file.

Click the button in the Layout Editor when you have the prototype frame laid out to your satisfaction.

This will bring up the Create Batch Report dialog where you can select the Quicktime tab (at right). When you select the Create button, a QuickTime movie will be generated with a frame for each sample in the current group. Movies will obey iteration settings.