Agilent Technologies NovoCyte Quanteon Operator's Manual page 79

Running Samples
In the example in this guide, setting the FSC-H threshold as 100,000 ensures the
NOTE
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
Cell Types
Cell lines, larger than 20 µ m in
cell diameter
Cells lines, smaller than 20 µ m
in cell diameter
Fixed or un-fixed, freshly
isolated cells (leukocytes,
spleen cells, thymocytes)
Platelets
Bacteria
Table 8
Standard Microsphere Diameter
0.1 µ m
0.2 µ m
0.5 µ m
2 µ m
>10 µ m
To detect microsphere with 0.1 µ m diameter, the photodetector gain of SSC needs
NOTE
to be increased. Please refer to
for instructions and more details on adjusting the photodetector gain.
NovoCyte Quanteon Flow Cytometer Operator's Guide
Recommended Threshold for Different Cell Types
FSC-H Threshold
300,000~1,000,000
100,000~300,000
50,000~200,000
5,000~10,000
1,000~10,000
Reference Threshold for Different Size of Standard Polystyrene Microspheres
Remark
Use linear or logarithmic scale for FSC
and SSC
Use linear or logarithmic scale for FSC
and SSC
Use linear or logarithmic scale for FSC
and SSC
Use logarithmic scale for FSC and SSC
Use logarithmic scale for FSC and SSC.
Recommend to use SSC-H > 5,000 as
the second threshold.
Threshold
SSC-H 700 (increase the gain of SSC to improve the
resolution)
SSC-H 1,000
FSC-H 500
FSC-H 50,000
FSC-H 500,000
Parameter Settings in NovoExpress Software Guide
79