AutoGate uses counting beads method to calculate Absolute Cell Count.
The formula applied is A/B*C*D where;
- A is the fcs event count of the target gate for the calculation (step 4 below)
- B is the fcs event count of the beads gate (step 2 below)
- C is the # of beads per ul provided in AutoGate setup (step 1 below)
- D is the # ul used to re-suspend cells provided in AutoGate setup (step 1 below)
1. In this method, user is expected to specify # beads per ul & # ul used to re-suspend cells as part of the setup process, as below
![](https://cytogenie.org/wp-content/uploads/2022/06/1.Input_Wizard-1024x266.png)
2. After completing the setup, the Beads gate need to be created on the gating side.
![](https://cytogenie.org/wp-content/uploads/2022/06/2.Create_Beads_Gate-1024x388.png)
3. Define the Beads gate for Absolute Cell Counting by right clicking on the beads gate and choosing Use “Beads” as bead count for absolute cell count
![](https://cytogenie.org/wp-content/uploads/2022/06/3.Define_Beads_Gate-1024x487.png)
4. Upon creating the gates for the samples, click View Statistics… from the toolbar to view the Cell count results
![](https://cytogenie.org/wp-content/uploads/2022/06/4.View_Statistics_Table-1024x215.png)
5. In the GatingTreeTable window, click Configure columns to add the Cell Count column
![](https://cytogenie.org/wp-content/uploads/2022/06/5.Configure_Columns-1024x129.png)
6. It opens up the Define columns for statistics table window. Under Select statistics section, move Absolute Cell Count to Use these box
![](https://cytogenie.org/wp-content/uploads/2022/06/6.Add_Column.png)
7. Absolute Cell Count is shown in the GatingTreeTable for all the gates.
![](https://cytogenie.org/wp-content/uploads/2022/06/7_Absolute_Cell_Count-1024x508.png)
8. Absolute cell count is also shown in the toolip on hovering over the gate