In the course of this example, we will:
•Set up the concentration calculator using the time of acquisition and the flow rate.
•Obtain a one time result.
(Note that the following sections build upon steps 1 through 7 in the Setting up the Beads Gates chapter)
1.Select the Tools→Calculate→Concentration Calculator command (Figure T22.2).
The Concentration Calculator dialog appears again as shown in Figure T22.2. The following steps all refer to Figure T22.4.
2.Select the File to analyze, in this case Monocytes.002, from the drop-down list.
3.Select Monocytes as the Gate to analyze, from the drop-down list.
4.Select the Time radio button to specify the Volume Calculation type.
The start and end times embedded in the FCS data file appear by default, and the Total Time is calculated automatically. You can change the start or end time manually, or enter a specific Total Time directly.
5.Input the Flow Rate used when acquiring the sample. In this case, since the data was collected on a cytometer that used a flow rate of 60 μl/min, we will use a value of 0.06 mL/min.
6.Click the Calculate button.
The Results section now displays the volume of the sample analyzed and the concentration of cells in the Monocytes gate
(Figure T22.4).
Figure T22.4 Concentration Calculator Dialog Using Time and Flow Rate
7.Click Cancel to close the Concentration Calculator dialog.
In the next example, we will calculate the concentration using volume ($VOL) keyword.