Is there a Pot of Gold at the End of the Spectrum?

Lannigan, Joanne

doi:10.1002/cyto.a.24186

Cited by 4 publications

(3 citation statements)

References 19 publications

(21 reference statements)

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“…To improve reagent, sample resource and processing efficiency, we explored panel design on the Cytek Aurora spectral cytometry platform. The advantages of spectral cytometry [5], spectral unmixing [6], and guidance on spectral cytometry panel design [7] have been reported elsewhere. We aimed for a 25‐biomarker immune monitoring panel, using OMIP‐042 [8] as a starting point while referencing major immunophenotypes described by the Human Immune Phenotyping Consortium [1], OMIP‐034 [9], and OMIP‐056 [10].…”

Section: Introductionmentioning

confidence: 99%

Analytical performance of a 25‐marker spectral cytometry immune monitoring assay in peripheral blood

Jensen

Wnek

2020

Cytometry Pt A

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Herein, we describe the development and analytical performance characteristics of a spectral flow cytometry assay for longitudinal immune monitoring biomarker applications in human whole blood and/or peripheral blood mononuclear cells (PBMCs). This 25 immune biomarker panel and robust gating strategy, developed in normal healthy volunteers, resolves memory, polarization, and activation markers on T, B, and NK cells as well as myeloid subpopulations including monocytes, dendritic cells, neutrophils, basophils, and myeloid‐derived suppressor cells (MDSCs). Three associated fluorescence‐minus‐multiple (FMM) designs are proposed as gating controls. To our knowledge, this is the first report to investigate intra‐run precision, post collection whole blood stability, and the impact of PBMC processing in the context of spectral cytometry. We achieved high intra‐run sample precision (<20% CV) for >95% of all gated immune cell populations analyzed across biospecimen types. Additionally, we explored the application of FlowSOM analysis and resulting impact on assay precision metrics. We observed biomarker stability in blood out to 24 h (86%), 48 h (83%), and 72 h (76%) while highlighting select markers (i.e., CXCR3) and rare cell subsets (i.e., naïve Tregs, plasmacytoid DCs, MDSCs) affected by storage and/or PBMC manipulation. Interrogation of sample type is imperative to coherent application of immune monitoring assays. Overall based on analytical performance, this 25‐biomarker spectral cytometry assay is sufficiently robust for implementation in translational research settings.

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Section: Introductionmentioning

confidence: 99%

Analytical performance of a 25‐marker spectral cytometry immune monitoring assay in peripheral blood

Jensen

Wnek

2020

Cytometry Pt A

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“…Recent advances in flow cytometers have enabled the collection of very narrow bands of light across the entire spectrum resulting from the excitation of multiple laser wavelengths. By doing so, the identification of a "spectral fingerprint" that is unique to each unique fluorophore becomes a way to better differentiate one fluorophore from another, even if they only differ slightly [1]. This new approach has become to be known as full-spectrum flow cytometry (FSFC) and this Special Issue is dedicated to highlighting some of the advancing applications that this technology has recently enabled.…”

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confidence: 99%

“…The number of publications continues to increase, with well over 900 publications to date. The value of the FS approach has been documented in several publications [1,11,14] and likewise so has the development of detailed protocols. [15,16] Researchers are only beginning to scratch the surface of what is possible with FSFC, and as more unique and functional fluorescent probes become available, the greater impact this technology will have on scientific discovery.…”

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confidence: 99%