Expanding NIST Calibration of Fluorescent Microspheres for Flow Cytometry to More Fluorescence Channels and Smaller Particles

Paul DeRose; Linhua Tian; Elzafir Elsheikh; Aaron Urbas; Yu-Zhong Zhang; Lili Wang

doi:10.3390/ma13184111

Expanding NIST Calibration of Fluorescent Microspheres for Flow Cytometry to More Fluorescence Channels and Smaller Particles

Materials (Basel). 2020 Sep 16;13(18):4111. doi: 10.3390/ma13184111.

Authors

Paul DeRose¹, Linhua Tian¹, Elzafir Elsheikh¹, Aaron Urbas², Yu-Zhong Zhang³, Lili Wang¹

Affiliations

¹ Biosystems and Biomaterials Division, National Institute of Standards and Technology (NIST), Gaithersburg, MD 20899, USA.
² Chemical Sciences Division, National Institute of Standards and Technology (NIST), Gaithersburg, MD 20899, USA.
³ Protein and Cell Analysis, Thermo Fisher Scientific, Eugene, OR 97402, USA.

Abstract

The National Institute of Standards and Technology (NIST), the National Institutes of Health (NIH) and other industry stakeholders have been working together to enable fluorescence intensities of flow cytometer calibration beads to be assigned quantitative equivalent reference fluorophore (ERF) values with high accuracy and precision. The ultimate goal of this effort is to accurately quantify the number of antibodies bound to individual living cells. The expansion of this effort to assign ERF values to more than 50 fluorescence channels and particles with diameters ranging from 10 μm down to 80 nm is reported here.

Keywords: NIST; beads; calibration; cytometer; flow cytometry; fluorescence channels; fluorophores; microspheres; nanospheres; reference materials; standards.