Flowjo manual compensation

1) by double-clicking the compensation badge from the workspace window (any compensated sample has this badge next to the sample in the left hand column down the workspace.) 2) by clicking the “View Matrix ” button in the main Compensation window. Matrix Editor Window. FlowJo software is used for the analysis of flow cytometry data. You can use FlowJo to analyze all of your flow cytometry data - regardless of the cytometer used to collect your data files. This tutorial will introduce you to FlowJo and to the 6 steps involved in analyzing a basic immunophenotyping experiment. 1. Load data into the Workspace. 2. Steps involved in compensation by FlowJo: Define positive and negative gates for each fluorescence channel requiring compensation. Open the compensation window; drag the positive and negative populations into the appropriate boxes or use the Compensation Wizard for extra flexibility. Compute and save the matrix.

The first gate works the best for calculating compensation because the MFI (shown as the dashed vertical line in FlowJo in Figure 2) is the highest. In all the other gates, there are cells that are brighter than the MFI used to calculate the compensation, so those gates have essentially created a control that breaks the rules of a "good. FlowJo's manual on compensation. Detailed overview of the compensation theory underpinning our calculations. If these techniques do not get you to your desired compensation, or if you have further questions or concerns you can always email FlowJo's technical support team: techsupport@www.doorway.ru 2. Manual Compensation. During the early years of multicolor flow cytometry, where most researchers were using fluorochromes, manual compensation was the vogue method to correct for the spillover of a fluorochrome into secondary detectors.

Starting with FlowJo version , you can easily change the compensation matrix values. When you create a new compensation matrix, FlowJo no longer automatically saves it to a file. You can edit the compensation matrices directly from FlowJo, using the Platform- Compensate Sample- Edit/Save Matrix command (from the Platform - Compensate Sample menu, you can also Remove Compensation). This page will help acquaint you with the various ways to interact with Compensation Matrices in FlowJo. For your convenience, this document is broken into chapters. Note, Compensation should always be preceded by scaling your data appropriately. Poorly scaled data will effect downstream compensation, so get into the habit of adjusting transforms through the “T” button in FlowJo’s graph window, prior to opening the compensation wizard for single stained control assignment. Compensation. Compensation has undergone a major redesign for FlowJo Version 10! Compensation in flow cytometry is the process of correcting for fluorescence spillover emissions. The detectors, or channels, in the instrument are designed to detect a very specific range of emissions. However, the fluorophores used in flow cytometry do not adhere to the exact range of emission detected by the instrument.