Apply compensation and scaling to all files in experiment.Choose the plot style foe the new plots.Select the output directory for the new experiment.Select the FACSDiva™ experiment to modify.Create a new experiment or modify an existing experiment.Simply right-click on a sample that contains the desired population hierarchy and gating, and from that dialogue, click on “Export to FACSDiva™.” A new window will open with the following options: When you have created the gating structure in FlowJo v10.6+ needed for repeating your experiment, you can modify an existing Diva experiment (.XML file) that and import that file into FACSDiva™. Whether you would like to import compensation and scaling onlyĪfter making these decisions, you will then need to click the “Import from FACSDiva™” button to bring the information into FlowJo.An FCS file with the scaling and compensation that you would like applied.The Global Worksheet with the gates that you are importing.The experiment using the file navigator.Once the Import from FACSDiva™ dialogue has been opened, you will be asked to choose: Next, right-click on any of loaded samples and select “Import from FACSDiva™” to open the Import from FACSDiva™ dialogue. With FlowJo v10.6 open, load your FCS files from the experiment by dropping the files into the FlowJo Workspace or from the Add Samples dialogue. Importing FACSDiva™ Experiments into FlowJo Choosing Directory Export will allow for the direct import into FlowJo, as the Zip archive will need to be unzipped first. XML Choosing Zip File Export will export a Zip archive file that contains your experiment. You will be given the option to export as a Directory or a Zip File.Ĭhoosing Directory Export will export a folder with your experiment. XML) by either right-clicking on the experiment and selecting Export –> Experiments from the dialogue or by choosing File –> Export –> Experiment on the FACSDiva™ ribbon. Once you have finished collecting your flow cytometry data on a BD cytometer running FACSDiva™, you will need to export your experiment file (FACSDiva™ experiments have a file extension of. Additionally, you can now create FACSDiva™ experiments in FlowJo for experiment reiterations and precise cell sorting gates. This means that gating information and population hierarchies don’t need to be lost in the handoff. Now, with FlowJo v10.6 and newer, FACSDiva™ experiments can be imported into FlowJo and FlowJo can export FACSDiva™ templates. Until now, FCS files were the only data from FACSDiva™ that could be imported into FlowJo. When you modify the matrix, the numbers are converted to give an estimation of the amount (percentage) of spillover into the other parameter.Since 1994, Flow Cytometrists have been using FlowJo to analyze data collected on BD cytometers to create statistics and figures pivotal to publications on immune subsets. In version 4.5 and after, the matrix can be modified right in FlowJo using the "Edit/Save Matrix" menu option. In very rough terms, the 0.1803 corresponds to approximately 18% spillover (PE-%FITC).Ĭlick here for instructions on modifying the matrix. Thus, 0.1803 is the spillover coefficient between FITC and PE, or the amount of FITC signal in the PE channel for a FITC-stained cell 0.0091 is the amount of the PE signal in the FITC channel for a PE-stained cell. The rows are the spectra of each channel the columns are the contributions of each fluor into the channels. The subsequent lines are the spillover coefficients, corresponding to each of the channels named. The names must match exactly use the names that appear in the $PnN keywords of the FCS file. ![]() Channels that are not in this list are not compensated channels in this list that are not in the sample are ignored. The second line has the names of the fluorescence channels that are being compensated. The first line has the title of the compensation matrix. ![]() If you save the matrix to a file (using the "Edit/Save Matrix" option), the file looks like this: CompMatrix When the "Compute" button in the compensation matrix dialog window is clicked, FlowJo generates a compensation matrix.
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