The relationship between mean channel selection and the calculated coefficient of variation

Area of Science:

• Flow Cytometry
• Analytical Chemistry
• Biophysics

Background:

• Coefficients of variation (CV) are standard metrics for assessing linear flow cytometry (FCM) system performance.
• The accuracy of calculated CV relies on the relationship between data channel width and the standard deviation (S.D.) of measured distributions.
• Existing methods for CV calculation can be unreliable due to channelization effects.

Purpose of the Study:

• To investigate the impact of channel width on the accuracy of calculated CV in linear flow cytometry.
• To determine the relationship between S.D., channel width, and calculated CV.
• To provide guidance on optimizing CV estimation for FCM system performance evaluation.

Main Methods:

• Analysis of calculated CV using the quotient of S.D. and mean value.
• Modeling the dependence of calculated CV on the ratio of S.D. to channel width.
• Simulating FCM data distributions across varying channel widths.

Main Results:

• Calculated CV is approximately 20% larger than true CV when channel width equals S.D.
• CV calculations become unreliable when S.D. is half the channel width.
• Small changes in mean value cause large CV variations when S.D. is narrower than channel width.

Conclusions:

• The accuracy of calculated CV is critically dependent on the interplay between S.D. and channel width.
• Mean channel selection must consider these limitations for reliable FCM system performance evaluation.
• Improving CV estimation accuracy in linear FCM is achievable by positioning distributions in higher-numbered channels.