Flow cytometry and related technologies are used in a wide variety of laboratory settings—research (immunology, cell biology, cancer), clinical testing and diagnosis, veterinary, drug discovery and development, botany, microbial testing for food safety, environmental and marine biology. No matter the application or laboratory testing environment, every laboratory generating flow cytometry data should be focused on generating robust, reproducible and comparable data. But how to accomplish this objective is not straightforward. Thus, several international initiatives have been created in order to provide cytometrists tools to reach this goal in their laboratories.
The Clinical and Laboratory Standards Institute (CLSI) published the guidance document, H62 - validation of assays performed by flow cytometry in October 2021. This comprehensive document addresses all aspects of conducting flow cytometry measurements in regulated as well as non-regulated settings and brings together information regarding best laboratory practices ranging from uncrating the instrument to reporting final results.
The National Institute of Standards and Technology (NIST) Flow Cytometry Standards Consortium was launched in February, 2021. The Consortium is part of the broader advanced therapy program at NIST. The purpose of this consortium is to provide a neutral forum for stakeholders in the biotechnology and healthcare sectors, as well as government agencies, to identify and address common measurement challenges, exchange ideas and jointly accelerate the development of standards and reference materials for quantitative flow cytometry. The consortium expects to develop measurement solutions, standards and best practices for flow cytometry that will enable more accurate measurements and improved reproducibility and comparability of measurement results.
ISEV-ISAC-ISTH EV Flow Cytometry Working Group (EV FC WG) published the MIFlowCyt-EV guidelines for single EV flow cytometry in 2020 and is completing a compendium of procedures and considerations for single EV flow cytometry for publication in 2022. MIFlowCyt-EV is a guidelines framework for the reporting of single EV flow cytometry methods and results. It covers pre-analytical steps (including sample collection, storage, and processing), instrumentation (including qualification and calibration) and assays (including sample staining, essential controls, gating, and reportable metrics). The manuscript includes a suggested “checklist” format that complements the existing general MIFlowCyt guidelines for flow cytometry measurements.
This presentation will raise awareness about best practices for quantitative cytometry by providing an introduction to these initiatives. In addition the presentation will discuss how to be involved in on-going and future initiatives.