Quantitative flow and image cytometry measurements are routinely used in research and clinical laboratories to measure the number of cells and the pattern of specific application/disease biomarkers on the surfaces and cytoplasm of cells of interests taken from cell cultures, the blood, lymph nodes, or bone marrow. The accurate measurement of the numbers of these biomarkers provides important information about cell properties, lineage, clonality, differentiation stage, and activation status. Thus, the reliability of these measurements has a significant impact on our understanding of cell states and disease diagnosis, prognosis, and on recommended therapies. Currently, a commonly accepted method for quality control of multiplexed flow and image cytometry measurements does not exist. This includes factors such as instrument linearity, response function, dynamic range, and sensitivity. Nor do calibration and quantification standards exist for the measurement of multiple biological biomarkers. This lack of standards and controls for flow and image cytometry hinders the progress of biological and medical research and healthcare delivery. Thus, it is critical to develop quantitative measurement methodologies and protocols for improving the accuracy and comparability of quantitative cytometry measurements across different instrument platforms. In addition to the standards effort, we are developing quantitative flow and image cytometry assays for studying intracellular signal transduction pathways associated with T- and B-cell activation for profiling the efficacy of drugs and other therapies, cytotoxicity assays for nanomaterial evaluation, and multiplexed assays for stem cell characterization.
Fluorescence calibration; Equivalent reference fluorophores (ERF); Antibodies bound per cell; Biological control; Measurement procedure; Biological applications;