Validation of cell‐based fluorescence assays: Practice guidelines from the ICSH and ICCS – part III – analytical issues

Davis, Bruce H.; Dasgupta, Amar; Kussick, Steven J.; Han, Jin‐Yeong; Estrellado, Annalee

doi:10.1002/cyto.b.21106

Cited by 98 publications

(113 citation statements)

References 68 publications

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“…GATING STRATEGY In adopting a "fit for purpose" gating strategy for identifying potentially rare PC events it is important to use a gating strategy which minimizes subjectivity and maximizes reproducibility (1). Inicial gates to identify PCs and discriminate them from all other BM cells are drawn generously enough to include as many PCs events as possible, allowing for variants that express lower CD38, CD45 and/or CD138 to be included, while excluding contaminating lymphocytes, doublets and other cellular debris (Fig.…”

Section: Flow Cytometry Data Analysis and Plasma Cellmentioning

confidence: 99%

Consensus guidelines on plasma cell myeloma minimal residual disease analysis and reporting

Arroz

Came

Lin

et al. 2015

Cytometry Part B Clinical

148

176

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Background: Major heterogeneity between laboratories in flow cytometry (FC) minimal residual disease (MRD) testing in multiple myeloma (MM) must be overcome. Cytometry societies such as the International Clinical Cytometry Society and the European Society for Clinical Cell Analysis recognize a strong need to establish minimally acceptable requirements and recommendations to perform such complex testing.Methods: A group of 11 flow cytometrists currently performing FC testing in MM using different instrumentation, panel designs ( 6-color) and analysis software compared the procedures between their respective laboratories and reviewed the literature to propose a consensus guideline on flow-MRD analysis and reporting in MM.Results/Conclusion: Consensus guidelines support i) the use of minimum of five initial gating parameters (CD38, CD138, CD45, forward, and sideward light scatter) within the same aliquot for accurate identification of the total plasma cell compartment; ii) the analysis of potentially aberrant phenotypic markers and to report the antigen expression pattern on neoplastic plasma cells as being reduced, normal or increased, when compared to a normal reference plasma cell immunophenotype (obtained using the same instrument and parameters); and iii) the percentage of total bone marrow plasma cells plus the percentages of both normal and neoplastic plasma cells within the total bone marrow plasma cell compartment, and over total bone marrow cells. Consensus guidelines on minimal current and future MRD analyses should target a lower limit of detection of 0.001%, and ideally a limit of quantification of 0.001%, which requires at least 3 3 10 6 and 5 3 10 6 bone marrow cells to be measured, respectively. V C 2015 International Clinical Cytometry Society

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Section: Flow Cytometry Data Analysis and Plasma Cellmentioning

confidence: 99%

Consensus guidelines on plasma cell myeloma minimal residual disease analysis and reporting

Arroz

Came

Lin

et al. 2015

Cytometry Part B Clinical

148

176

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“…With proper calibration through it is possible to express fluorescence intensity in stoichiometric units called molecules of equivalent soluble fluorochrome (MESF) that reflect the concentrations of the fluorescent conjugates and the receptors they stain (62). Protocols have been developed for the validation of cellbased fluorescence assays that can be adapted for measurement of microvesicles (63).…”

Section: Microvesicle Size Calibration and Gatingmentioning

confidence: 99%

Measurement of microvesicle levels in human blood using flow cytometry

Chandler

2016

Cytometry Part B Clinical

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Microvesicles are fragments of cells released when the cells are activated, injured, or apoptotic. Analysis of microvesicle levels in blood has the potential to shed new light on the pathophysiology of many diseases. Flow cytometry is currently the only method that can simultaneously separate true lipid microvesicles from other microparticles in blood, determine the cell of origin and other microvesicle characteristics, and handle large numbers of clinical samples with a reasonable effort, but expanded use of flow cytometric measurement of microvesicle levels as a clinical and research tool requires improved, standardized assays. The goal of this review is to aid investigators in applying current best practices to microvesicle measurements. First pre-analytical factors are evaluated and data summarized for anticoagulant effects, sample transport and centrifugation. Next flow cytometer optimization is reviewed including interference from background in buffers and reagents, accurate microvesicle counting, swarm interference, and other types of coincidence errors, size calibration, and detection limits using light scattering, impedance and fluorescence. Finally current progress on method standardization is discussed and a summary of current best practices provided. V C 2016 Clinical Cytometry Society

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“…Flow cytometry was performed on fresh venous blood (BD Biosciences), using a FACS Canto II 6-colour flow cytometer, daily calibrated [43] with Calibrite beads (Fitc, Pe, PerCP and APC) and Compbeads (Pe-Cy7 and APC-Cy7; Becton Dickinson, San Jose, CA, USA). For identification of circulating MDSCs: FITC-anti-Lineage 1 antibodies (CD3, CD14, CD16, CD19, CD20, CD56), PE-anti-CD11b, PercP-anti-CD33, PE-Cy7-anti-HLA-DR, APC-anti-CD15 and APC-Cy7-anti-CD14 (BD Bioscience, San Diego, CA, USA).…”

Section: Antibodies and Flow Cytometric Analysismentioning

confidence: 99%

Peripheral myeloid-derived suppressor and t-regulatory pd-1 positive cells predict response to neoadjuvant short-course radiotherapy in rectal cancer patients

Cardone¹,

Napolitano²,

Rega³

et al. 2016

European Journal of Surgical Oncology (EJSO)

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Validation of cell‐based fluorescence assays: Practice guidelines from the ICSH and ICCS – part III – analytical issues

Cited by 98 publications

References 68 publications

Consensus guidelines on plasma cell myeloma minimal residual disease analysis and reporting

Consensus guidelines on plasma cell myeloma minimal residual disease analysis and reporting

Measurement of microvesicle levels in human blood using flow cytometry

Peripheral myeloid-derived suppressor and t-regulatory pd-1 positive cells predict response to neoadjuvant short-course radiotherapy in rectal cancer patients

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