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flowdex

Extract Fluorescence Distribution Data from FCS Files and Recalculate to Events per Volume

Description

Extract fluorescence distribution data from any bivariate distribution within a previously defined flow cytometry gating strategy and recalculate the fluorescence distribution to events per volume unit.

Additionally, any number of class- and numerical variables describing the dataset can be generated by providing a structured character string in the Sample-ID field of each individual sample at the time of data acquisition. At the time of reading in the fcs files, this structured character string is translated into class- and numerical variables, using a dictionary to translate any abbreviations into its long form.

Finally, the fluorescence-distribution data that were (possibly) re-calculated to events per volume unit can be visualised and exported to file, along with data denoting the overall events per volume unit in each sample in each gate.

To (meaningfully) use flowdex, the fcs files have to contain volumetric measurement data denoting the acquired sample volume.

flowdex is leaning heavily on the packages flowCoreand flowWorkspace for data import, generation of gating sets etc.

To sum it up, package flowdex can:

  • Extract fluorescence distribution data from FCM files and recalculate them to events per volume unit,
  • Generate variables describing the dataset by using a structured character string in the sample-Id field of the sample at the time of data acquisition, and
  • Visualise fluorescence distributions and export them to file.

Advantage

Having ones hand directly on the rawdata of fluorescence distributions is not something the average desktop GUI of FCM-machines is providing easily or willingly. Package flowdex was designed to alleviate that problem.

Additionally, if the FCM-machine used for data acquisition does have a volumetric measurement module and the fcs files contain volumetric data, it is possible to re-calculate any extracted fluorescence distribution to events per volume unit.

In other words, the y-axis of any fluorescence distribution can be displayed in a fixed scale, i.e. events per volume unit. This enables the comparison of numbers on a fixed scale across samples, groups or experiments, instead of being confined to compare percentages of sub-groups of an overall population.

Installation

Install release version from CRAN via

install.packages("flowdex") 

Or download from github:

library(devtools)
install_github(repo="bpollner/flowdex", ref="main")

Go to https://bpollner.github.io/flowdex/ to learn how to use flowdex.

flowdex's People

Contributors

bpollner avatar

Watchers

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flowdex's Issues

Have filename suffix when exporting the fdmat-obj. and the raw data

Like in the plotgates function, it would be nice to have the possibility to add a custom filename suffix to the name when exporting (also automatically) the fdmat-object and the file containing the raw data.
Because so far, these names are always the same when using flowdexit, and the user has to manually change / modify the name so that previous files do not get overwritten.

Message displayed when plotting gates

When plotting gates using plotgates(), the following message is displayed:
"Coordinate system already present. Adding new coordinate system, which will replace the existing one."
This is caused by adding
ggcyto::ggcyto_par_set(limits="instrument")
to the plot object.
A possible solution to avoid this (harmless) message might be to extract the machine limits manually from a flow frame, then create the plot object with these limits already.
As time is a factor at the moment, I will leave this as it is for now.

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