Full spectrum flow cytometry in the clinical laboratory

Abstract

Contemporary full spectrum or “spectral” flow cytometry is a recently developed technology that allows for high‐dimensional flow cytometric analyses of cells and particles in suspension. This single‐cell technology has gained popularity in research settings because it can conservatively detect 35 or more antigens simultaneously in a single‐tube assay format. Recently, spectral flow cytometry has obtained regulatory approval for use as an in vitro diagnostic device in China and Europe, enabling use of this technology in some clinical flow cytometry laboratories. The purpose of this review is to describe the basic principles of conventional and spectral flow cytometry, contrasting these two technologies. To illustrate the analytic power of spectral flow cytometry, we provide an example of spectral flow cytometry data analyses and the use of a machine learning algorithm to harvest the vast amount of information contained within large spectral flow cytometry datasets. Finally, we discuss the advantages of spectral flow cytometry adoption in clinical laboratories and preliminary studies comparing the performance of this technology relative to conventional flow cytometers that are currently used in clinical laboratory environments.

Phenotypic Analysis of Hematopoietic Stem and Progenitor Cell Populations in Acute Myeloid Leukemia Based on Spectral Flow Cytometry, a 20-Color Panel, and Unsupervised Learning Algorithms

“…The advantages of spectral cytometers, as highlighted in previous research [ 29 ], include their cost-effectiveness and reduced sample material requirements compared to traditional cytometers. Additionally, the use of extended multicolor panels, as opposed to the currently prevalent 8–13 color panels, enhances precision by enabling the assessment of a greater number of markers in a single tube.…”

Applications of Flow Cytometry in Drug Discovery and Translational Research

“…We anticipate that the maturation of several technologies and analysis approaches will contribute to the evolution of clinical flow cytometry, enabling an increased clinical impact in the coming years. Firstly, the maturation of flow cytometric hardware, including mass cytometry and spectral flow cytometry, will greatly enable flexibility in panel design and expand the number of parameters that can be evaluated in a single sample [116]. Secondly, the evolution of multi-omic and AI/ML-driven data analysis approaches will enable automated, multiparametric evaluation of cell populations [117,118], which may ultimately serve to reduce the variability conferred by manual definition of cell populations and gating.…”

Biomarker-based human and animal sperm phenotyping: the good, the bad and the ugly

“…New image-based flow cytometers also come equipped with image-based spectral sorting. Spectral flow cytometry could aid andrologists in deriving more information from a single sample and offers an improved framework for cellular characterization and analysis [ 238 , 239 ], which could enable a new generation of sperm sorting techniques, such as sorting for superior morphology sperm. This could be a bridge to help improve human IVF, which currently highly relies on ICSI as opposed to traditional IVF.…”