76 BD LSR II User’s Guide
Criteria for DNA Experiments
To support DNA experiments, a flow cytometer must provide:
• ability to resolve histogram peaks (populations)
• linearity of the DNA fluorescence signal
• ability to distinguish singlets from aggregates
While obtaining good peak resolution for DNA also depends on proper sample
preparation, optimization of instrument optics and fluidics is critical. A flow
cytometer’s ability to resolve peaks can be assessed by measuring the CV of a
reference particle: the lower the CV, the better the resolution.
Linearity is also critical for DNA experiments. To assess the linearity of DNA
data, the pulse-area signal is used to measure the amount of DNA fluorescence
detected from cells or nuclei. On an instrument with good linearity, the doublet
peak should be located at twice the mean channel of the singlet peak (Figure 4-1).
Figure 4-1 Area signal and DNA fluorescence
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LSR2.book Page 76 Tuesday, April 25, 2006 3:34 PM
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