By labeling samples with a numerical keyword that uniquely identifies the separate files, you can both:
- combine populations from different files into one FCS data file, and
- create a new parameter from any such keyword.
Combining populations in FlowJo® is easy using concatenation. Simply select a set of populations you’d like to combine, and run Export/Concatenate. Find this feature on the File tab:
In the following example, we show a set of titration samples with different antibody concentrations, denoted (in μg/ml) by the “Concentration” keyword:
Within the Advanced Options section of the Concatenation dialog box, the keyword we wish to parameterize is added as an additional parameter, and the new concatenated data file is constructed:
Now, when analyzing the concatenated data file, these different samples can easily be gated on and separated for investigation:
This feature can also be useful in conjunction with FlowJo’s tSNE plugin. The tSNE function helps researchers automatically cluster samples in two dimensions based on a much larger number of predefined parameters.
Because the tSNE plugin is non-deterministic, it is often more useful to run it on a concatenated set of samples. Having run tSNE on your fluorescence parameters of interest, you can then separate the individual samples to see how their clusters differ:
For more information regarding FlowJo’s Export/Concatenation platform, check out the following documentation page:
To discover more about the tSNE plugin, try this page:
If you still have questions or want to discuss your data in particular, we encourage you to write to FlowJo tech support at firstname.lastname@example.org.