Agilent Technologies NovoCyte Quanteon, Operator'S Manual, Page: 79 / 148

Running Samples
NovoCyte Quanteon Flow Cytometer Operator's Guide 79
NOTE
In the example in this guide, setting the FSC-H threshold as 100,000 ensures the
correct collection of the leukocyte signals, and effectively discards most of the
impurities. Refer to Table 7 as a starting point for setting the threshold for different
biological samples. Table 8 serves as a reference to determine which threshold
value to start from based on microsphere size. Some biological samples may
contain a significant amount of impurities or debris. In this case, setting a second
threshold with a fluorescent signal helps to improve the quality of the collected
data.
Table 7 Recommended Threshold for Different Cell Types
Cell Types FSC-H Threshold Remark
Cell lines, larger than 20 µm in
cell diameter 300,000~1,000,000 Use linear or logarithmic scale for FSC
and SSC
Cells lines, smaller than 20 µm
in cell diameter 100,000~300,000 Use linear or logarithmic scale for FSC
and SSC
Fixed or un-fixed, freshly
isolated cells (leukocytes,
spleen cells, thymocytes) 50,000~200,000 Use linear or logarithmic scale for FSC
and SSC
Platelets 5,000~10,000 Use logarithmic scale for FSC and SSC
Bacteria 1,000~10,000 Use logarithmic scale for FSC and SSC.
Recommend to use SSC-H > 5,000 as
the second threshold.
Table 8 Reference Threshold for Different Size of Standard Polystyrene Microspheres
Standard Microsphere Diameter Threshold
0.1 µm SSC-H 700 (increase the gain of SSC to improve the
resolution)
0.2 µm SSC-H 1,000
0.5 µm FSC-H 500
2 µm FSC-H 50,000
>10 µm FSC-H 500,000
NOTE
To detect microsphere with 0.1 µm diameter, the photodetector gain of SSC needs
to be increased. Please refer to
Parameter Settings in NovoExpress Software Guide
for instructions and more details on adjusting the photodetector gain.