Calculating Sample Volume And Concentration - Beckman Coulter CytoFLEX Instructions For Use Manual

Calculating Sample Volume and Concentration

The CytoFLEX flow cytometer supports the calculation of the sample concentration based on the
volume consumed and/or based on the known concentration of reference beads.
NOTE
If necessary, calibrate the sample uptake rate (see
Replacement/Adjustment
•
Select the cells/μL(V) checkbox to calculate concentration directly.
NOTE
• If using reference beads to calculate the concentration, select the cells/μL(B) checkbox and select
the gated Beads Population. Enter the total number of reference beads as the Beads Count, as well
as the sample volume in total. The software automatically calculates the original sample
concentration based on the input values. (You can also enter the reference bead concentration
directly in the beads count field and set the sample volume as 1.)
To obtain accurate calculations, throughout the data acquisition process, ensure that:
• The sample concentration is 2 x 10
• Samples are thoroughly mixed before loading and that they exhibit no apparent subsidence
throughout the testing process.
• The detection rate is maintained at less than 10,000 events/second throughout the sampling
process. When the detection rate does not exceed the stated event rate, running at medium to
high acquisition speeds are considered more accurate.
• A constant sampling rate is maintained when recording data.
• You acquire at least 10 μL of sampling volume.
B49006AS
Procedures) prior to collecting data for volumetric analysis:
The direct calculation of concentration can be affected by several conditions such as, the
sample's viscosity and sample mixing. Uncalibrated sample volume uptake rates may lead to
erroneous results.
Calculating Sample Volume and Concentration
Calibrating the Sample Flow Rate
4 7
-10 units/mL.
8
Data Review
in CHAPTER 12,
8-7