Cytometry Part A最新文献_第9页

Best Practices for Equitable Training in Flow Cytometry Shared Resource Laboratories 流式细胞术共享资源实验室公平培训的最佳实践。

IF 2.1 4区生物学

Cytometry Part A Pub Date : 2025-10-03 DOI: 10.1002/cyto.a.24965

Jane Srivastava

引用次数: 0

Volume 107A, Number 9, September 2025 Cover Image 107A卷，第9号，2025年9月封面图片

IF 2.1 4区生物学

Cytometry Part A Pub Date : 2025-09-29 DOI: 10.1002/cyto.a.24873

引用次数: 0

OMIP-117: 40-Parameter/37-Color Spectral Cytometry Panel for Robust Immunoprofiling of Human Lymphoid Subsets in Cancer Patients 40参数/37色光谱细胞仪检测癌症患者淋巴亚群的免疫谱图

IF 2.1 4区生物学

Cytometry Part A Pub Date : 2025-09-24 DOI: 10.1002/cyto.a.24962

Ondrej Venglar, Eva Radova, Lucie Broskevicova, Roman Hajek, Tomas Jelinek

{"title":"OMIP-117: 40-Parameter/37-Color Spectral Cytometry Panel for Robust Immunoprofiling of Human Lymphoid Subsets in Cancer Patients","authors":"Ondrej Venglar, Eva Radova, Lucie Broskevicova, Roman Hajek, Tomas Jelinek","doi":"10.1002/cyto.a.24962","DOIUrl":"10.1002/cyto.a.24962","url":null,"abstract":"<p>The analysis of immune cell compartments in cancer patients is crucial to predict treatment efficacy and relapse. We introduce a robust 40-parameter, 37-channel spectral cytometry panel designed to profile human lymphoid subsets and CAR-T cell expansion, with the capability to assess exhaustion status by profiling immune checkpoints and activating receptors in cancer patients. Developed for the 5-laser Cytek Aurora, the panel optimizes fluorophore selection and uses three pairs of mutually exclusive markers assigned to a single fluorescent parameter to simplify setup and ensure robust data, adopting a conservative design choice to keep similarity indices below 0.85; though higher overlaps can still yield high-quality data when best practices are applied. The panel enables detailed analysis of well-defined lymphoid subsets using a conventional gating strategy, as well as detection of unconventional subsets with variable expression patterns by unsupervised algorithm-based analysis. The effectiveness of the panel is demonstrated through a dataset simulating the progression of multiple myeloma, from pre-malignant disease to a highly aggressive stage.</p>","PeriodicalId":11068,"journal":{"name":"Cytometry Part A","volume":"107 10","pages":"641-648"},"PeriodicalIF":2.1,"publicationDate":"2025-09-24","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/cyto.a.24962","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145130257","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Monitoring Treatment-Associated Receptor Expression in Multiple Sclerosis Using a Newly Developed Panel for Spectral Flow Cytometry 使用新开发的光谱流式细胞术面板监测多发性硬化治疗相关受体的表达。

IF 2.1 4区生物学

Cytometry Part A Pub Date : 2025-09-23 DOI: 10.1002/cyto.a.24963

Katharina Robichon, Jennifer Taylor, Imogen Milner, Kathryn Elizabeth Hally, Anne Camille La Flamme

{"title":"Monitoring Treatment-Associated Receptor Expression in Multiple Sclerosis Using a Newly Developed Panel for Spectral Flow Cytometry","authors":"Katharina Robichon, Jennifer Taylor, Imogen Milner, Kathryn Elizabeth Hally, Anne Camille La Flamme","doi":"10.1002/cyto.a.24963","DOIUrl":"10.1002/cyto.a.24963","url":null,"abstract":"<p>Multiple sclerosis (MS) is a chronic, immune-mediated autoimmune disease characterized by the infiltration of autoreactive T cells and other inflammatory immune cells from the periphery into the central nervous system. Currently, there is no cure for MS, and treatment consists of disease-modifying therapies (DMTs), most of which modify or delete specific immune cell populations. These populations express crucial MS treatment-associated receptors, which may be differentially expressed in each patient, and thus each drug may affect individuals differently. Here, we developed the first comprehensive 24-parameter flow cytometry immunophenotyping panel to evaluate treatment-associated receptor expression on the major MS-associated immune cells in whole blood. Analyzing whole blood samples from treatment-naïve individuals with MS using this panel, we demonstrated that expression levels of these receptors vary between individuals. Response to the chosen DMT treatment also differed across participants. When monitoring the receptor expression during the course of treatment, we detected an increased response to treatment when receptor expression was elevated at the start of treatment. This panel reliably detects these receptors in MS treatment-naïve participants and enables monitoring of their expression throughout treatment. This tool will enable deep interrogation of the immune receptors targeted by MS therapies and highlights that treatment-associated receptor expression levels might be used to predict or correlate with treatment response.</p>","PeriodicalId":11068,"journal":{"name":"Cytometry Part A","volume":"107 10","pages":"668-682"},"PeriodicalIF":2.1,"publicationDate":"2025-09-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/cyto.a.24963","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145124446","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Novel Spectral High-Dimensional Flow Cytometry Assay for Combinatorial MHC Class I Tetramer Staining and Deep Antigen-Specific CD8+ T Cell Phenotyping 新型光谱高维流式细胞术用于组合MHC I类四聚体染色和深度抗原特异性CD8+ T细胞表型。

IF 2.1 4区生物学

Cytometry Part A Pub Date : 2025-09-18 DOI: 10.1002/cyto.a.24959

William Pratcher, Chikara Takahashi, Maria Lorenzo, Alberto Robert, Jiun Chiun Chang, Leesun Kim, Daniel Haensel, Martine Darwish, Mahesh Yadav, William E. O'Gorman, Thomas Liechti

{"title":"Novel Spectral High-Dimensional Flow Cytometry Assay for Combinatorial MHC Class I Tetramer Staining and Deep Antigen-Specific CD8+ T Cell Phenotyping","authors":"William Pratcher, Chikara Takahashi, Maria Lorenzo, Alberto Robert, Jiun Chiun Chang, Leesun Kim, Daniel Haensel, Martine Darwish, Mahesh Yadav, William E. O'Gorman, Thomas Liechti","doi":"10.1002/cyto.a.24959","DOIUrl":"10.1002/cyto.a.24959","url":null,"abstract":"<div>\u0000 \u0000 <p>Cytotoxic CD8+ T cells eliminate virus-infected or cancer cells, thus playing a pivotal role in anti-viral and anti-cancer immunity. Tetramer reagents, which consist of fluorochrome-labeled streptavidin coupled with peptide-loaded MHC I molecules, enable the detection of antigen-specific CD8+ T cells using flow cytometry. The development of tetramer reagents has been instrumental for our understanding of antigen-specific CD8+ T cells and their roles in immune responses. More recently, combinatorial tetramer staining protocols have enabled the simultaneous detection and monitoring of multiple specificities and concomitant pathogen-dependent CD8+ T cell dynamics. However, these methods are either based on mass cytometry, preventing the isolation of antigen-specific CD8+ T cells for downstream investigation, or have provided a less comprehensive picture of the phenotypic characteristics of antigen-specific CD8+ T cells when based on flow cytometry. Here we describe the development of a combinatorial tetramer staining protocol in combination with high-dimensional CD8+ T cell immunophenotyping in the context of virus-specific CD8+ T cells leveraging spectral flow cytometry. Our assay enables the simultaneous measurement of 15 different CD8+ T cell specificities and includes an additional 18 markers to define the phenotypic and functional characteristics of antigen-specific CD8+ T cells. We describe our assay optimization strategies, with the goal of improving marker and tetramer resolution while eliminating sources of background noise. Finally, we apply this method to reveal the phenotypic heterogeneity of virus-specific CD8+ T cells against common viral pathogens in healthy individuals.</p>\u0000 </div>","PeriodicalId":11068,"journal":{"name":"Cytometry Part A","volume":"107 9","pages":"614-628"},"PeriodicalIF":2.1,"publicationDate":"2025-09-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145079827","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Impact of Panel Size, Fluorochrome Selection, and Unmixing Algorithms on Ultra-High Parameter Flow Cytometry Analysis 面板尺寸、荧光染料选择和解混算法对超高参数流式细胞术分析的影响。

IF 2.1 4区生物学

Cytometry Part A Pub Date : 2025-09-17 DOI: 10.1002/cyto.a.24960

Debajit Bhowmick, Timothy P. Bushnell

{"title":"Impact of Panel Size, Fluorochrome Selection, and Unmixing Algorithms on Ultra-High Parameter Flow Cytometry Analysis","authors":"Debajit Bhowmick, Timothy P. Bushnell","doi":"10.1002/cyto.a.24960","DOIUrl":"10.1002/cyto.a.24960","url":null,"abstract":"<div>\u0000 \u0000 <p>The expansion of full spectral flow cytometry enabled us to run ultra-high-dimensional panels of up to 50 fluorochromes, offering unprecedented in-depth immunophenotyping. However, this advancement introduces significant analytical challenges, particularly in unmixing accuracy, population spread, and panel design. This study evaluates the impact of various unmixing algorithms on biological interpretation using different OMIP datasets. We demonstrate that algorithmic discrepancies can lead to loss of resolution, population misidentification, and incorrect interpretation of the biological information. Through comparative analysis and the use of measures like the Median Mismatch Index (MMI), Spillover Spread Matrix (SSM) and robust Standard Deviation (rSD), we highlight the limitations of current tools and propose strategies for optimized use of single stain, predicting the unmixing accuracy for a set of fluorochromes, and cares that need to be taken for correct data interpretation in high-parameter cytometry. We also showed the present version of SSM may not be suitable to predict the spillover spread for ultra-large panels.</p>\u0000 </div>","PeriodicalId":11068,"journal":{"name":"Cytometry Part A","volume":"107 10","pages":"683-694"},"PeriodicalIF":2.1,"publicationDate":"2025-09-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145074541","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Development of a 29-Color Single-Tube Full Spectrum Flow Cytometry Assay for the Detection of Measurable Residual Disease and Leukemic Stem Cells in Acute Myeloid Leukemia 29色单管全光谱流式细胞术检测急性髓系白血病中可测量的残留疾病和白血病干细胞的建立。

IF 2.1 4区生物学

Cytometry Part A Pub Date : 2025-09-12 DOI: 10.1002/cyto.a.24958

Zhong Zhang, Miriam Wilhelm, Ines Sieber, Hartmut Döhner, Michaela Feuring

{"title":"Development of a 29-Color Single-Tube Full Spectrum Flow Cytometry Assay for the Detection of Measurable Residual Disease and Leukemic Stem Cells in Acute Myeloid Leukemia","authors":"Zhong Zhang, Miriam Wilhelm, Ines Sieber, Hartmut Döhner, Michaela Feuring","doi":"10.1002/cyto.a.24958","DOIUrl":"10.1002/cyto.a.24958","url":null,"abstract":"<p>Multiparametric flow cytometry (MFC) is widely used to detect measurable residual disease (MRD) in acute myeloid leukemia (AML). However, conventional flow assays require multiple tubes, with an additional tube for leukemia stem cell (LSC) analysis and lack hemodilution evaluation. Spectral flow cytometry (SFC) can overcome the limitation of flow channels and has the potential for multifunctional design using a single tube. We developed a 29-color single-tube assay that adheres to the recommendations of the European Leukemia Network Flow-MRD Working Party and incorporates the simultaneous evaluation of MRD, LSC, and hemodilution. The Complexity Index of the assay was calculated at 9.08. Through limit dilution experiments using the KG-1α AML cell line, we determined the limit of blank (LOB), limit of detection (LOD), and limit of quantification (LOQ) for four leukemia-associated immunophenotypes (LAIP). The assay easily achieved the minimum sensitivity requirement for MRD detection ≤ 0.1% with minimal intra- and interassay variations. Background signals for 24 LAIPs and 10 LSC immunophenotypes were evaluated in eight healthy bone marrow (BM) samples. The single-tube SFC assay was compared with the five-tube conventional assay by analyzing 20 AML BM samples, demonstrating high concordance. To assess hemodilution, markers to detect established parameters, including immature granulocytes, neutrophils, mast cells, and plasma cells, were included. In summary, we provide a versatile single-tube 29-color SFC-based MRD assay that minimizes cell requirements, integrates LSC evaluation, and assesses hemodilution. This assay has the potential to improve the reliability and simplicity of MRD detection.</p>","PeriodicalId":11068,"journal":{"name":"Cytometry Part A","volume":"107 9","pages":"597-613"},"PeriodicalIF":2.1,"publicationDate":"2025-09-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/cyto.a.24958","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145039442","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Volume 107A, Number 8, August 2025 Cover Image 107A卷，8号，2025年8月封面图片

IF 2.1 4区生物学

Cytometry Part A Pub Date : 2025-09-09 DOI: 10.1002/cyto.a.24871

引用次数: 0

Per-Event Uncertainty Quantification for Flow Cytometry Using Calibration Beads 流式细胞术中使用校准珠的每事件不确定度定量。

IF 2.1 4区生物学

Cytometry Part A Pub Date : 2025-08-29 DOI: 10.1002/cyto.a.24954

Prajakta Bedekar, Megan A. Catterton, Matthew DiSalvo, Gregory A. Cooksey, Anthony J. Kearsley, Paul N. Patrone

{"title":"Per-Event Uncertainty Quantification for Flow Cytometry Using Calibration Beads","authors":"Prajakta Bedekar, Megan A. Catterton, Matthew DiSalvo, Gregory A. Cooksey, Anthony J. Kearsley, Paul N. Patrone","doi":"10.1002/cyto.a.24954","DOIUrl":"10.1002/cyto.a.24954","url":null,"abstract":"<div>\u0000 \u0000 <p>Flow cytometry measurements are widely used in diagnostics and medical decision making. Incomplete understanding of sources of measurement uncertainty can make it difficult to distinguish autofluorescence and background sources from signals of interest. Moreover, established methods for modeling uncertainty overlook the fact that the apparent distribution of measurements is a convolution of the inherent population variability (e.g., associated with calibration beads or cells) and instrument-induced effects. Such issues make it difficult, for example, to identify signals from small objects such as extracellular vesicles. To overcome such limitations, we formulate an explicit probabilistic measurement model that accounts for volume and labeling variation, background signals, and fluorescence shot noise. Using raw data from routine per-event calibration measurements, we use this model to separate the aforementioned sources of uncertainty and demonstrate how such information can be used to facilitate decision making and instrument characterization.</p>\u0000 </div>","PeriodicalId":11068,"journal":{"name":"Cytometry Part A","volume":"107 9","pages":"587-596"},"PeriodicalIF":2.1,"publicationDate":"2025-08-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144946169","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

CytoNormPy Enables a Fast and Scalable Removal of Batch Effects in Cytometry Datasets CytoNormPy能够快速和可扩展地去除细胞计数数据集中的批处理效果。

IF 2.1 4区生物学

Cytometry Part A Pub Date : 2025-08-29 DOI: 10.1002/cyto.a.24953

Tarik Exner, Nicolaj Hackert, Luca Leomazzi, Sofie Van Gassen, Yvan Saeys, Hanns-Martin Lorenz, Ricardo Grieshaber-Bouyer

引用次数: 0