Flow cytometry results

Data output: FCS file

Each sample acquired by the instrument forms a data file that is called an FCS file (Flow Cytometry Standard file). It contains all the raw data for each individual cell. All quantitative data for the parameters of light scatter and fluorescence are individually stored in that file. The file may contain data for up to 10X106 cells.

Data analysis

FCS files can be analyzed by programs that recognize specifically this file format or they may be converted to text files and analyzed by mathematical programs such as "Matlab". Some of the specific FCS recognizing programs are used for acquisition as well as for analysis while others are capable of analysis only.

Our Flow Unit uses "FACSDiva" for the PC-controlled machines for both acquisition and analysis. Additional programs are used exclusively for analysis and are installed on PC off-line analysis computer.
The programs available are: "FlowJo", "FCS-Express", "Modfit" and "Multicycle". Data files can be transferred to these computers for analysis by file sharing, mail or by using the "Bioimg" storage system of the institute. Please note that work on these computers is free of charge but reservations have to be made in order to ensure your analysis time.

Imaging Flow Cytometry: Analysis Results

Data output: RIF file

Each sample acquired by the ImageStreamX forms a data file that is called a RIF file (Raw Image File). It contains the images for each individual cell, in the acquired channels. The usual file contains data for 104

cells, but usually not more than 105 cells are acquired.

Data analysis

The images acquired are analyzed by the IDEAS software, installed on the ImageStreamX analysis station. The software can be downloaded for free from the Amnis website (www.amnis.com), following a short registration. Data files can be transferred by file sharing or by using the "Bioimg" storage system of the institute.

Support with Data analysis

Support of the FACS staff is given to enable use of the programs so that results can be analyzed and their significance understood in biological terms.
In order to receive help with analysis - the "help needed" box must be checked and help has to be coordinated with the staff before reserving the analysis period.

Only basic support can be promised for the use of the various analysis programs and it is up to the users to master their full capabilities and follow their upgrades.