Fluorescence drift detection as a novel QC procedure for DNA cell-cycle analysis

Area of Science:

• Biotechnology
• Molecular Biology
• Genetics

Background:

• DNA cell-cycle analysis is crucial for understanding cell proliferation and ploidy.
• Fluorescence detection during data acquisition can be subject to drift, potentially compromising results.
• Existing methods may lack objectivity or reproducibility in detecting such artifacts.

Purpose of the Study:

• To develop an objective, quantitative, and reproducible method for detecting fluorescence drift in DNA cell-cycle data.
• To introduce quality control software, TruPloid, for identifying and quantifying fluorescence drift.
• To highlight the impact of fluorescence drift on DNA cell-cycle histogram analysis.

Main Methods:

• Development of quality control software named "TruPloid."
• Analysis of list-mode files using three distinct statistical tests.
• Quantification of fluorescence drift to assess its impact on data.

Main Results:

• Fluorescence drift can cause measurement artifacts such as high coefficients of variation.
• Drift can obscure small cell populations and create artificial peaks in histograms.
• Forty percent of 50 archived list-mode files exhibited detectable fluorescence drift.

Conclusions:

• Fluorescence drift is a significant source of artifacts in DNA cell-cycle histogram analysis.
• The TruPloid software provides a reliable method for detecting and quantifying fluorescence drift.
• Implementing objective detection methods is essential for ensuring the accuracy of DNA cell-cycle data.