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A simple and rapid method for determining the linearity of a flow cytometer amplification system
C B Bagwell1, D Baker, S Whetstone
1Maine Cytometry Research Institute, Portland 04102.
Cytometry
|November 1, 1989
Summary
This study presents a quick method to check flow cytometer amplifier linearity. Deviations from a straight line indicate nonlinearities, with correlation coefficients quantifying the degree of nonlinearity.
Area of Science:
- Biotechnology
- Analytical Chemistry
- Instrumentation
Background:
- Flow cytometry is a crucial technique for cell analysis.
- Accurate amplification is essential for reliable flow cytometry data.
- Nonlinear amplification can lead to significant data inaccuracies.
Purpose of the Study:
- To develop a simple and rapid method for assessing flow cytometer amplification linearity.
- To identify nonlinear components within flow cytometer systems.
- To provide a means for quantifying nonlinearity and enabling compensation.
Main Methods:
- Utilizing two bead or cell populations with slight fluorescence differences.
- Analyzing samples across increasing photomultiplier tube high-voltage settings.
- Plotting the mean difference versus mean position to assess linearity.
Main Results:
- Linear amplifiers produce a straight-line plot intersecting the origin.
- Deviations from linearity indicate the presence of nonlinear components.
- The correlation coefficient quantifies the degree of nonlinearity detected.
Conclusions:
- The described method offers a straightforward approach to evaluate flow cytometer linearity.
- This technique can detect certain types of nonlinearities in the amplification system.
- A method for compensating amplifier nonlinearity is also presented.