CYTO 2026 Scientific Tutorial: Key Steps for Assay Standardization across Cytometer Platforms

Key Steps for Assay Standardization across Cytometer Platforms

Presenters:
Lili Wang, PhD, Senior Scientist, National Institute of Standards and Technology (NIST)
Huan-Yu (Ray) Chen, PhD, post-doctoral researcher, National Tsing Hua University
Paul DeRose, PhD, physical/analytical chemist, NIST
Yu-Fen (Andrea) Wang, MS, cofounder and CEO, AHEAD Medicine Corporation

Abstract:
This tutorial will address the critical need for comparable and standardized data in flow cytometry, a challenge that the NIST Flow Cytometry Standards Consortium (FCSC) Working Group 2 (WG2) tackled with its first Interlaboratory Study (ILS). The study focused on a 10-marker, 8-color T/B/Monocyte/Natural Killer (TBMNK) cell assay—to establish measurement confidence across diverse instruments. The ILS involved 21 sites and 42 different cytometer platforms (including conventional, spectral, and imaging types) from 7 instrument companies, utilizing cryopreserved peripheral blood mononuclear cells (PBMC) and TruCytes TBMNK synthetic cells serving as QC samples.

In this tutorial, we will detail the study's framework by walking through the four SOPs developed for the ILS:

WG2 SOP-01: Designated for instrument set up, calibration and characterization
WG2 SOP-02: Preparation and acquisition of three compensation matrices with one cell-based and two bead-based matrices
WG2 SOP-03: Preparation and acquisition of study samples as well as various assay controls, including fluorescence minus one (FMO) controls
WG2 SOP-04: Data analysis and result reporting 

Finally, we will present the study data quality control criteria and the key summary of data analysis from manual analysis by participants and centralized automated analysis pipeline. The tutorial will conclude with practical instructions on how to navigate the interactive site that provides all the results from this comprehensive ILS study.

Keywords: Fundamental Cytometry Concepts, Instrument Calibration, Instrument Standardization, NIST, FCS Consortium, Assay Standardization

CMLE Credit: 1.5