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Material Measurement Laboratory

Biosystems and Biomaterials Division

FCSC Working Group 1

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Schematic demonstrating, in the upper half, how comparing the fluorescence spectra of reference fluorophore solutions to the fluorescence spectra of calibration beads allows, in the lower half, calibrations made using different calibration beads to become comparable measurements through the equivalent number of reference fluorophores (ERF) value.

Comparative measurement of reference fluorophore solutions and fluorescent calibration beads allows comparison of calibrations performed using different beads through the assigned value of Equivalent Number Reference Fluorophores (ERF).

ERF-Based Instrument Calibration and Standardization

Working Group 1 (WG1) is developing standards including reference materials, reference data, and reference methods for the calibration of flow cytometers and the standardization of flow cytometry data.  Activities in progress include:

  1. A measurement service available to Consortium members for assigning the Equivalent Number of Reference Fluorophores (ERF) to calibration microspheres along with associated uncertainties  This service supports the ultimate goal to establish an absolute fluorescence intensity scale that is instrument independent.
  2. An interlaboratory study for cross-calibration with four different ERF bead sets for ERF assignment of unknowns  Data submission is complete with contributions from 17 Consortium members across 22 sites and 47 different instrument platforms including imaging, spectral, and conventional flow cytometers.  The goals of the study are:
    • Determine the variability in instrument calibration across bead sets and instrument types.
    • Compare different approaches to calibration data analysis.

Return to NIST Flow Cytometry Standards Consortium main page

Bioscience, Bioeconomy, Biological data, Biomanufacturing, Biometrology, Cell measurements, Gene delivery systems, Health, Cell and gene therapy, Clinical diagnostics, Cancer, Precision medicine, Regenerative medicine and advanced therapy, Flow cytometry, Metrology, Amount of substance, Standards, Reference data and Reference materials

Contacts

Working Group 1 Technical Lead

Consortium Principal Investigator

Created November 29, 2024, Updated March 10, 2025