Evaluation of Voltration Approaches for Optimal Data Acquisition in Flow Cytometry

Abstract

Flow cytometry is a technology widely used to analyze biophysical and biochemical characteristics of cells and small particles quickly with high sensitivity at the single cell level. Due to its high sensitivity, broad dynamic range of quantification, robust reproducibility, and acquisition of multiplexed data at single cell level, flow cytometry has seen appreciable utilities in scientific research, biomedical industry and clinic. However, despite its increasing popularity, the quality of flow cytometry data can vary substantially, either between experiments, individuals, or laboratories. This prevalent problem can be attributed to multiple sources such as instrument standardization practices, reagent standardization, equipment configuration, user operation, quality of data acquisition, and data processing. Overcoming these challenges has been a major goal for the flow cytometry community, however not all sources of flow cytometric experimental error are equally obvious to even experienced users. In this thesis work, I systemically evaluated how the standardization method used for assessing instrument electronic noise and configuring operation voltages can contribute to observed experimental variances. Through side-by-side comparisons between these eight methods, I provide comprehensive operational suggestions to preserve both data quality and real-world practicality.