Standardization, Calibration, and Control in Flow Cytometry

Wang, Lili; Hoffman, R. A.

doi:10.1002/cpcy.14

Cited by 64 publications

(70 citation statements)

References 87 publications

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“…Set up and optimize the flow cytometer's voltages and compensation for data acquisition using standardized methods as described by Wang and Hoffman, (2017). Set up and optimize the flow cytometer's voltages and compensation for data acquisition using standardized methods as described by Wang and Hoffman, (2017).…”

Section: Methodsmentioning

confidence: 99%

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Monitoring of Measurable Residual Disease in Multiple Myeloma by Multiparametric Flow Cytometry

Soh

Wallace

2019

CP Cytometry

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Recent interest in high‐sensitivity multiple myeloma (MM) measurable residual disease (MRD) testing is a direct consequence of the high‐quality responses achieved using novel therapeutic agents and better treatment strategies. Traditional diagnostic measures such as immunohistochemistry and morphology have detection sensitivities of only 10−2 to 10−3, which do not reliably predict progression‐free survival (PFS) or overall survival (OS) after these treatments. Contemporary monitoring of MM MRD has switched to more sensitive platforms such as allele‐specific oligonucleotide quantitative polymerase chain reaction (ASO‐qPCR), next‐generation sequencing (NGS), and multiparametric flow cytometry (MFC). Though both ASO‐qPCR and NGS have excellent detection sensitivities (10−5 to 10−6), both technologies have lower applicability when compared to MFC. Conventional MFC can easily reach a detection sensitivity of 10−4 and when optimized can achieve a sensitivity of 10−5 to 10−6. Current consensus guidelines require a minimum of 2 million and recommend 5 million events be acquired to reach a minimum sensitivity of 10−5. As conventional immunophenotyping protocols are unable to attain these numbers, alternative MFC staining procedures are required. This article describes two high‐sensitivity MFC approaches that can be used for MM MRD testing. © 2019 by John Wiley & Sons, Inc.

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Section: Methodsmentioning

confidence: 99%

“…15. Set up and optimize the flow cytometer's voltages and compensation for data acquisition using standardized methods as described by Wang and Hoffman, (2017). Adjust threshold setting based on the forward scatter light characteristics to include hematogones while judiciously avoiding the recording of unwanted background noise.…”

Section: Of 29mentioning

confidence: 99%

Monitoring of Measurable Residual Disease in Multiple Myeloma by Multiparametric Flow Cytometry

Soh

Wallace

2019

CP Cytometry

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“…As a result, critical methods of instrument standardization and calibration (27)(28)(29)(30)(31), panel and experimental design (32)(33)(34)(35)(36), sample preparation and controls (37)(38)(39), spillover correction (40), and data presentation and publication (41,42) are emphasized in various seminal publications. As a result, critical methods of instrument standardization and calibration (27)(28)(29)(30)(31), panel and experimental design (32)(33)(34)(35)(36), sample preparation and controls (37)(38)(39), spillover correction (40), and data presentation and publication (41,42) are emphasized in various seminal publications.…”

Section: Existing Flow Cytometry Resources and Guidelines On Preparinmentioning

confidence: 99%

MiSet RFC Standards: Defining a Universal Minimum Set of Standards Required for Reproducibility and Rigor in Research Flow Cytometry Experiments

et al. 2019

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Poor adherence to best practices, insufficient training, and pressure to produce data quickly may lead to publications of suboptimal biomedical research flow cytometry data, which contributes to the body of irreproducible research findings. In addition, documentation of compliance with best flow cytometry practices for submission, visualization, and publication of flow cytometry data is currently endorsed by very few scientific journals, which is particularly concerning as numerous peer-reviewed flow cytometry publications emphasize instrumentation, experimental design, and data analysis as important sources of variability. Guidelines and resources for adequate reporting, annotation and deposition of flow cytometry experiments are provided by MIFlowCyt and the FlowRepository database, and comprehensive expert recommendations covering principles and techniques of field-specific flow cytometry applications have been published. To facilitate the integration of quality-defining parameters into manuscript and grant submission and publication requirements across biomedical fields that rely on the use of flow-cytometry-based techniques, a single comprehensive yet easily and universally applicable document is needed. To produce such a list of gold-standard parameters that assess whether a research flow cytometry experiment has been planned, conducted, interpreted, and reported at the highest standard, a new initiative defining the minimum set of standards a robust and rigorous research flow experiment must fulfill (MiSet RFC Standards) was proposed at CYTO 2019. MiSet RFC Standards will integrate and simplify existing resources to provide a universal benchmark a flow cytometry experiment can easily be measured against. The goal of MiSET RFC Standards is its integration into peerreview and publication procedures through partnership with stakeholders, journals and publishers in biomedical and translational research. This article introduces the aims and anticipated timeline and discusses strategies for interdisciplinary consensus and implementation. A single-resource broadly applicable guideline will harmonize standards across different fields of biomedical research and lead to publication of more robust research findings.

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“…Additional resources for using a flow cytometer can be found here: Hoffman (2005) and Wang & Hoffman (2017).…”

Section: Data Acquisitionmentioning

confidence: 99%

Quantification of Cellular Proteostasis in Live Cells by Fluorogenic Assay Using the AgHalo Sensor

Fares

Zhang

2018

CP Chemical Biology

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Proper cellular proteostasis is essential to cellular fitness and viability. Exogenous stress conditions compromise proteostasis and cause aggregation of cellular proteins. We have developed a fluorogenic sensor (AgHalo) to quantify stress‐induced proteostasis deficiency. The AgHalo sensor uses a destabilized HaloTag variant to represent aggregation‐prone cellular proteins and is equipped with a series of fluorogenic probes that exhibit a fluorescence increase when the sensor forms either soluble oligomers or insoluble aggregates. Herein, we present protocols that describe how the AgHalo sensor can be employed to visualize and quantify proteome stress in live cells using a direct fluorescence read‐out and visualization with a fluorescence microplate reader and a microscope. Additionally, protocols for using the AgHalo sensor in combination with fluorogenic probes and commercially available HaloTag probes to enable two‐color imaging experiments are described. These protocols will enable use of the AgHalo sensor to visualize and quantify proteostasis in live cells, a task that is difficult to accomplish using previous, always‐fluorescent methods. © 2018 by John Wiley & Sons, Inc.

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Standardization, Calibration, and Control in Flow Cytometry

Cited by 64 publications

References 87 publications

Monitoring of Measurable Residual Disease in Multiple Myeloma by Multiparametric Flow Cytometry

Monitoring of Measurable Residual Disease in Multiple Myeloma by Multiparametric Flow Cytometry

MiSet RFC Standards: Defining a Universal Minimum Set of Standards Required for Reproducibility and Rigor in Research Flow Cytometry Experiments

Quantification of Cellular Proteostasis in Live Cells by Fluorogenic Assay Using the AgHalo Sensor

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