Mercurial > repos > immport-devteam > cross_sample
diff src/README @ 1:7eab80f86779 draft
add FLOCK
author | immport-devteam |
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date | Mon, 27 Feb 2017 13:26:09 -0500 |
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--- /dev/null Thu Jan 01 00:00:00 1970 +0000 +++ b/src/README Mon Feb 27 13:26:09 2017 -0500 @@ -0,0 +1,139 @@ +This package contains R code for converting and transforming a binary +FCS file, using the FCSTrans software, and the C code for running the +flock1 and flock2 population identification software. + +src - contains the C code for flock1, flock2 and cent_adjust +bin - contains the compiled C code for flock1, flock2 and cent_adjust, + plus the R code for using the FCSTrans algorithm for file + conversion and data transformation. +doc - documentation for FCSTrans and FLOCK algorithms +example - sample data and output from FCSTrans and FLOCK + +To run this software sucessfully the code assumes the software was installed +in the /usr/local/flock directory and that R, plus the Bioconductor flowCore +module has been installed. If you use the ipconvert.sh shell script, you +may need to adjust the location of the RScript executable and the location +of the FCSTrans.R code by editing the shell script. + +############################################################################# +# Overview +############################################################################# +ImmPort-FLOCK + +FLOCK (FLOw Clustering without K), an automated population discovery tool for +multidimensional FCM data was designed to specifically take into account the +unique feature of FCM data and produce objective segregation of cell +populations. + +FLOCK parameter settings can be customized by defining Bins and Density +Threshold. The number of bins is an integer specifying the number of +equal-sized regions the data will be partitioned into on each axis. Increasing +the number of bins increases the sensitivity to detect rare populations but +may also result in single populations being divided. Density Threshold is the +cut-off value to separate the dense regions from background. It is a floating +point number that helps define population centers; increasing the threshold +may help separate major populations but could cause the algorithm to overlook +rare populations. + +############################################################################# +# Compiling C code +############################################################################# +cd bin +cc -o flock1 ../src/flock1.c ../src/find_connected.c -lm +cc -o flock2 ../src/flock2.c -lm +cc -o cent_adjust ../src/cent_adjust.c -lm + +############################################################################# +# FCSTrans +############################################################################# +A shell script named ipconvert.sh is included that runs the FCSTrans R +code for converting and transforming a binary FCS file. The output consists +of one text file containing the transformed channel intensity values and +another file containing a list of the FCS parameters. + +cd bin +/usr/local/flock/bin/ipconvert.sh ../example/data/FCS2.fcs +/usr/local/flock/bin/ipconvert.sh ../example/data/FCS3.fcs + +############################################################################# +# Running flock1 or flock2 +############################################################################# +Running the FLOCK1 and FLOCK2 algorithms generate 8 output files that have +generic file names. For this reason, it is recommended that one output +directory be created for one input file to the program. + +cd example/output/FCS2 +/usr/local/flock/bin/flock1 ../../data/FCS2.txt + +cd example/output/FCS3 +/usr/local/flock/bin/flock2 ../../data/FCS3.txt + +Files created: MFI.txt, percentage.txt, population_id.txt, profile.txt, + flock_results.txt, coordinates.txt, population_center.txt + and percentage.txt + +Usage Information for FLOCK1 +---------------------------------------------------------------------------- +basic mode: flock1 fcs.txt +advanced_ mode: flock1 fcs.txt num_bin density_index max_num_pop + +Usage Information for FLOCK2 +---------------------------------------------------------------------------- +basic mode: flock data_file +advanced mode 0 (specify maximum # of pops): flock data_file max_num_pop +advanced mode 1 (without # of pops): flock data_file num_bin density_index +advanced mode 2 (specify # of pops): flock data_file num_bin density_index + number_of_pop +advanced mode 3 (specify both # of pops): flock data_file num_bin density_index + number_of_pop max_num_pop + +FLOCK Output Files +---------------------------------------------------------------------------- +coordinates.txt: + Output is the intensity values for each marker and event + +flock_results.txt: + A combination of the input file, event identifiers and population + identifiers. + +MFI.txt: + Provides the mean fluorescence intensity for each population for each + marker/parameter + +population_id.txt: + Contains population identifiers (i.e, values from [1 to n] where n is + the population assigned to the corresponding events in the input data + file, one identifier per row.) + +population_center.txt: + Contains the centroid coordinates for each identified population + +percentage.txt: + Includes the population identifiers and percentage of events within that + population (relative to the whole data file) + +profile.txt: + Displays an expression profile, where the approximate expression level + for each marker is assigned a numeric value from 1-4, for each identified + population + +fcs_properties.txt: + Contains the number of events, number of populations, and number of + markers, as well as the algorithm parameters used in the analysis + +############################################################################# +# Running cent_adjust +############################################################################# +Running the cent_adjust algorithm generates 4 output files that have +generic file names. For this reason, it is recommened that one output +directory be created for one input file to the program. + +mkdir example/output/FCS2/cent_adjust +cd example/output/FCS2/cent_adjust +/usr/local/flock/bin/cent_adjust ../population_center.txt ../coordinates.txt + +Files created: MFI.txt, percentage.txt, population_id.txt and profile.txt + +Usage Information for cent_adjust +---------------------------------------------------------------------------- +basic mode: cent_adjust input_center input_data_file