If you would like the uncompensated always overlaid by default, check this box in the preferences. Overlay uncompensated – The matrix editor will allow you to overlay your uncompensated data on top of the compensated to view the difference. Proper compensation requires that the positive and negative populations be gated on similar cell types (i.e., that have the same autofluorescence when unstained). If you prefer to not see this confirmation box, uncheck the box here in the preferences. First of all, we will open the first sample (FITC Comp sample), in order to make a lymphocyte gate. These preferences can be very useful since you have the control to set the default values of whatever keyword you wish to automatically assign compensation single stain controls to the compensation group upon loading the files! Matrix EditorĬonfirm Edit – when you click the “Edit” button in the matrix editor, a confirmation box will open asking you to confirm that you wish to edit the matrix. The interface will open as shown below: Compensation Group Inclusion The Group Inclusion can be used to make files with certain file names get automatically included in the group when they are loaded into FlowJo. Some other common keywords are “TUBENAME” and “SampleID”. Select the Compensation button in the Preferences tool to open the Compensation preferences. If you prefer to use a different keyword, you can remove the default $FIL strings using the X icon next to them in this interface and then click the “Add” button to add whatever keywords you would like to use. This tool works for matrices created in FlowJo, as the single stain controls are needed to calculate several statistics from to produce each value in the SSM. Thus, any FCS files put into a workspace that contains the text “comp” or “unstained” in the $FIL, or filename, will be included in the compensation group in your workspace when loaded.
The default values set are “comp” and “unstained”. $FIL – is the FCS file keyword for the name of the data file written by the cytometer. The equation calculates the ratio of the DIFFERENCE in the secondary fluorescence over the DIFFERENCE in primary fluorescence (essentially, a slope).
#Compensation flowjo software
In fact, the brighter, the better Compensation controls allow the software to calculate how much spillover fluorescence occurs from that level of primary fluorescence. The compensation matrix is represented by the grid to the left of the sample name. Can my compensation control be too bright No.
The “Group Inclusion” can be used to make files with certain file names get automatically included in the group when they are loaded into FlowJo. Create new, apply old, or compare different matrices. With the proper controls, the matrix creation process should be. The workflow was constructed in this manner to discourage users from editing spillover values while creating the matrix. Select the Compensation button in the Preferences tool to open the Compensation preferences. CHAPTER 3 The Matrix Editor The process of creating a compensation matrix and the process of editing a compensation are considered two separate processes in version 10. Set your default preferences for compensation
0 kommentar(er)
