Nicole So, Sara Monaghan, Katelynn Davis, Grant Bullock, Wendy Shallenberger, Ahmad Al-Attar
The objective of this 12-color/13-antibody single-tube panel is to assist in the diagnosis of T/NK-cell leukemias and lymphomas. In the clinical setting, the absence of a standardized T/NK-cell panel limits inter-laboratory data comparability, contributes to diagnostic variability, and results in redundant efforts across laboratories to design and validate panels for flow cytometry. Developing and promoting a panel that allows for rapid and fairly comprehensive labeling of surface T/NK-cell markers, including the anti-T-cell receptor β-chain constant region 1 (TRBC1) antibody, will streamline the workflow by establishing a standard immunophenotyping T/NK-cell panel for detecting neoplastic T/NK cells and lay the foundation for future COMIPs.
{"title":"COMIP-001: A clinical 13-antibody, 12-color flow cytometry panel for optimized immunophenotyping of T/NK cells in human leukemia and lymphoma diagnostics.","authors":"Nicole So, Sara Monaghan, Katelynn Davis, Grant Bullock, Wendy Shallenberger, Ahmad Al-Attar","doi":"10.1002/cytob.70015","DOIUrl":"https://doi.org/10.1002/cytob.70015","url":null,"abstract":"<p><p>The objective of this 12-color/13-antibody single-tube panel is to assist in the diagnosis of T/NK-cell leukemias and lymphomas. In the clinical setting, the absence of a standardized T/NK-cell panel limits inter-laboratory data comparability, contributes to diagnostic variability, and results in redundant efforts across laboratories to design and validate panels for flow cytometry. Developing and promoting a panel that allows for rapid and fairly comprehensive labeling of surface T/NK-cell markers, including the anti-T-cell receptor β-chain constant region 1 (TRBC1) antibody, will streamline the workflow by establishing a standard immunophenotyping T/NK-cell panel for detecting neoplastic T/NK cells and lay the foundation for future COMIPs.</p>","PeriodicalId":10883,"journal":{"name":"Cytometry Part B: Clinical Cytometry","volume":" ","pages":""},"PeriodicalIF":2.7,"publicationDate":"2026-02-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146141351","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
<p>[Color figure can be viewed at wileyonlinelibrary.com]</p><p>This issue of Clinical Cytometry B deals with several topics which are relevant in either the hematology and oncology discipline, mostly focusing on multiple myeloma and malignant lymphoma. Two of these studies reported data on standardization of flow cytometry methods which may be of great value in several clinical situations. It is a personal pleasure to comment here on these interesting manuscripts.</p><p>The first paper of this issue was written by Deutsch-Biedermann et al. (<span>2026</span>); it was related to the development of a one-tube flow cytometry panel for measurable residual disease (MRD) detection in multiple myeloma patients (MM). Recent data has clearly demonstrated that the achievement of MRD negativity after first-line therapy for MM is one of the strongest surrogate markers for both overall survival and progression-free survival and is now widely used as an endpoint in several clinical trials (Anderson et al., <span>2021</span>; Mateos et al., <span>2019</span>; Munshi et al., <span>2020</span>). It is also known that flow cytometry represents the leading technique for MRD status assessment in MM and related diseases (Devitt et al., <span>2025</span>). Furthermore, compliance with the International Myeloma working group (IMWG) Next Generation Flow (NGF) criteria for MRD negativity requires the absence of immunophenotypically abnormal and clonal plasma cells with a minimum sensitivity of 1 in 10<sup>5</sup> nucleated cells in bone marrow aspirates using the EuroFlow standard method or a validated equivalent method. Interestingly, in this manuscript, authors have proposed a novel MRD panel based on the use of all EuroFlow recommended immunophenotypic markers (cytoplasmic light chain kappa and lambda, CD19, CD27, CD38, CD45, CD56, CD81, CD117, CD138) other than the CD200 marker, which allows the inclusion of a total of 11 fluorochromes in one tube (Terra et al., <span>2025</span>). This protocol meets the quality specifications of the IMWG. This flow cytometry protocol showed higher sensitivity than the previously developed flow panel and showed concordant results with the EuroFlow NGF panel, thus sustaining the reliability and sensitivity of this approach for detecting MRD in MM patients.</p><p>In the paper by Groves et al. (<span>2026</span>) the authors provide guidance for performing minor modifications to validated antibody panels designed to ensure consistent and accurate results in the immunophenotyping of hematopoietic cells in several diseases. However, it is well known that in unforeseen situations, such as unique or unusual immunophenotypes or supply chain issues, ad hoc modifications to these panels are strongly necessary (D'Addio et al., <span>2025</span>; Kelleher et al., <span>2024</span>; Lanza et al., <span>2025</span>; Terra et al., <span>2025</span>; Tian et al., <span>2024</span>). This manuscript provides guidance for performing minor modificati
本期《临床细胞术B》涉及几个与血液学和肿瘤学相关的主题，主要关注多发性骨髓瘤和恶性淋巴瘤。其中两项研究报告了流式细胞术方法标准化的数据，这可能在几种临床情况下具有重要价值。在这里评论这些有趣的手稿是我个人的荣幸。本期第一篇论文由Deutsch-Biedermann et al.（2026）撰写；这与用于多发性骨髓瘤（MM）患者可测量残留疾病（MRD）检测的单管流式细胞仪面板的发展有关。最近的数据清楚地表明，MM一线治疗后MRD阴性是总生存期和无进展生存期的最强替代标志物之一，现在被广泛用作几个临床试验的终点（Anderson等人，2021；Mateos等人，2019；Munshi等人，2020）。众所周知，流式细胞术是评估MM及相关疾病的MRD状态的领先技术（Devitt et al., 2025）。此外，符合国际骨髓瘤工作组（IMWG）下一代流量（NGF） MRD阴性标准要求，使用EuroFlow标准方法或经过验证的等效方法，骨髓抽吸液中不存在免疫表型异常和克隆浆细胞，其最低敏感性为105个有核细胞中有1个。有趣的是，在这篇论文中，作者提出了一种新的MRD面板，基于使用除CD200标记之外的所有EuroFlow推荐的免疫表型标记（细胞质轻链kappa和lambda， CD19, CD27, CD38, CD45, CD56, CD81, CD117, CD138），它允许在一个管中包含总共11种荧光染料（Terra等人，2025）。该协议符合IMWG的质量规范。该流式细胞术方案显示出比先前开发的流式面板更高的灵敏度，并显示出与EuroFlow NGF面板一致的结果，从而维持了该方法检测MM患者MRD的可靠性和灵敏度。在Groves等人（2026）的论文中，作者为对经过验证的抗体板进行微小修改提供了指导，这些抗体板旨在确保几种疾病中造血细胞免疫分型结果的一致性和准确性。然而，众所周知，在不可预见的情况下，例如独特或不寻常的免疫表型或供应链问题，对这些面板进行特别修改是非常必要的（D'Addio等人，2025；Kelleher等人，2024；Lanza等人，2025；Terra等人，2025；Tian等人，2024）。本手稿提供了进行微小修改的指导，例如替换或添加一或两个抗体，同时保持检测的完整性。随着一些淋巴细胞增殖性疾病的免疫治疗的出现，在使用抗cd20和CD19治疗后，必须修改用于监测患者的抗体组。事实上，缺乏CD19和CD20表达的B细胞淋巴瘤，除非在实验室可用的面板中添加或替换另一种标记物，否则无法可靠地评估肿瘤细胞的谱系。本研究中提出的建议和最佳实践试图通过允许实验室在不影响检测性能的情况下调整抗体面板来优化患者护理。这可以通过评估对抗体结合、测定灵敏度、荧光补偿和总体测定性能的影响来实现。作者还强调了实验室医学主任在监督文件编制、审查和批准经修订的方案方面发挥的作用，其主要目的是减轻与这些修改相关的风险。在Chiriac等人（2026）的这篇文章中，研究了CD200表达作为ssamzary综合征（SS）的附加诊断标志物。虽然CD200在b细胞肿瘤中的表达已经得到了很好的证实，但在t细胞肿瘤中的表达却很少被研究。初步结果已报道结节性T滤泡辅助淋巴瘤（nTFHL）和外周T细胞淋巴瘤（PTCL）的免疫组化。目前的数据与恬厚和罗山（2022）的研究结果一致，与t -原淋巴细胞白血病（T-PLL）和非肿瘤CD4+ t细胞相比，循环ssamzary细胞中CD200的表达更高。此外，CD200表达的定量评估显示，nTFHL中肿瘤CD4+ T细胞与非肿瘤CD8+ T细胞的MFI比显著高于其他所有成熟T细胞肿瘤，尤其是PTCL、NOS和非肿瘤T细胞。总之，CD200表达评估似乎可以将SS与其他t细胞肿瘤区分开来，并提示CD200评估可用于评估PB在t细胞肿瘤中的影响（Bellesi等）。 , 2024;Wang等人，2025)。然而，基于这些结果，可能推测需要更大规模的队列研究来证实这些发现，并评估CD200表达作为TFHL的潜在治疗和预后指标。在Ecker等人（2026）的论文中，研究了抗CD38 VHH MoAb （JK36）在daratumumab治疗的多发性骨髓瘤（MM）患者中的诊断作用，以更好地评估接受CD38免疫治疗的患者骨髓瘤细胞的持久性。在过去的几年中，MM的治疗方案发生了巨大的变化，多种新型药物的可用性越来越高，包括moab，如daratumumab和isatuximab，这些药物大大改善了患者的预后（Rocchi等人，2024；Zannetti等人，2021）。实现最深层的反应，达到最小残留病（MRD）阴性并保持持续缓解是迈向MM治愈的重要步骤。考虑到Daratumumab最近在移植项目中被引入，关于其对造血干细胞（HSC）收集的影响的数据仍处于初步阶段（Horenstein等人，2020；Zannetti等人，2020）。初步数据显示，daratumumab不会对成功进行自体干细胞移植（ASCT）的可能性产生负面影响，ASCT仍然是有资格接受大剂量化疗的患者的治疗基石。然而，最近的研究似乎表明，基于daratumumab的诱导疗法可能与较低的动员和收集HSC的能力有关（Bigi等人，2024；Valentini等人，2025）。众所周知，治疗性抗体通常与传统的分析性单克隆抗体竞争，限制了研究被占领抗原表面表达的能力。在细胞水平上，CD38暴露后，抗原下调可能发生；此外，消失和再现的动力与许多因素有关，可能因情况而异（Chen et al., 2023; Malavasi et al., 2021）。天然存在于骆驼体内的重链抗体（vhh）的可变重结构域，由于其扩展的互补决定区3 (CDR3)，可以克服这些干扰。由于扩展的互补决定区3 (CDR3), CD38（VHH）抗体可以识别常规抗体无法识别的靶抗原空腔内的3d样表位（Bannas et al., 2017; Bruins et al., 2024）。在这篇细胞术手稿中，使用了由两个抗CD38 VHH片段组成的克隆JK36，其主要目的是允许靶向传统抗体无法接近的隐性CD38表位。将该诊断试验与daratumumab治疗（d-t）后MM骨髓样本中的常规抗cd38抗体（LS198） （CA）与未治疗(n)和未知治疗样本进行比较。共分析了111份样本。在未接受治疗的样本中，VHH和CA同样能很好地检测到CD38，但CD38只能在8%的CA d-t样本中检测到，而在91%的VHH中检测到。这导致可检测PC的数量总体显着减少，与VHH相比，CA检测不到PC的三个样本。此外，在52%的d-t样本中，CD138被减少/降解，其中88%的样本无法通过CA检测到CD38。本文还表明，VHH抗体评估了细胞表面CD38表达的减少，与n样品相比，d-t上CD38中位荧光强度（MFI）下降。有趣的是，在一个d-t样本中，两种不同的PC群体因CD38表达暗淡和明亮而不同，只有VHH可以检测到。综上所述，VHH在检测d-t MM患者的CD38表达方面更为可靠。这些数据可能对MM研究的几个领域感兴趣，包括造血干细胞移植和最小残留病（MRD）测量。此外，
{"title":"Issue Highlights—January 2026","authors":"Francesco Lanza","doi":"10.1002/cytob.70007","DOIUrl":"10.1002/cytob.70007","url":null,"abstract":"<p>[Color figure can be viewed at wileyonlinelibrary.com]</p><p>This issue of Clinical Cytometry B deals with several topics which are relevant in either the hematology and oncology discipline, mostly focusing on multiple myeloma and malignant lymphoma. Two of these studies reported data on standardization of flow cytometry methods which may be of great value in several clinical situations. It is a personal pleasure to comment here on these interesting manuscripts.</p><p>The first paper of this issue was written by Deutsch-Biedermann et al. (<span>2026</span>); it was related to the development of a one-tube flow cytometry panel for measurable residual disease (MRD) detection in multiple myeloma patients (MM). Recent data has clearly demonstrated that the achievement of MRD negativity after first-line therapy for MM is one of the strongest surrogate markers for both overall survival and progression-free survival and is now widely used as an endpoint in several clinical trials (Anderson et al., <span>2021</span>; Mateos et al., <span>2019</span>; Munshi et al., <span>2020</span>). It is also known that flow cytometry represents the leading technique for MRD status assessment in MM and related diseases (Devitt et al., <span>2025</span>). Furthermore, compliance with the International Myeloma working group (IMWG) Next Generation Flow (NGF) criteria for MRD negativity requires the absence of immunophenotypically abnormal and clonal plasma cells with a minimum sensitivity of 1 in 10<sup>5</sup> nucleated cells in bone marrow aspirates using the EuroFlow standard method or a validated equivalent method. Interestingly, in this manuscript, authors have proposed a novel MRD panel based on the use of all EuroFlow recommended immunophenotypic markers (cytoplasmic light chain kappa and lambda, CD19, CD27, CD38, CD45, CD56, CD81, CD117, CD138) other than the CD200 marker, which allows the inclusion of a total of 11 fluorochromes in one tube (Terra et al., <span>2025</span>). This protocol meets the quality specifications of the IMWG. This flow cytometry protocol showed higher sensitivity than the previously developed flow panel and showed concordant results with the EuroFlow NGF panel, thus sustaining the reliability and sensitivity of this approach for detecting MRD in MM patients.</p><p>In the paper by Groves et al. (<span>2026</span>) the authors provide guidance for performing minor modifications to validated antibody panels designed to ensure consistent and accurate results in the immunophenotyping of hematopoietic cells in several diseases. However, it is well known that in unforeseen situations, such as unique or unusual immunophenotypes or supply chain issues, ad hoc modifications to these panels are strongly necessary (D'Addio et al., <span>2025</span>; Kelleher et al., <span>2024</span>; Lanza et al., <span>2025</span>; Terra et al., <span>2025</span>; Tian et al., <span>2024</span>). This manuscript provides guidance for performing minor modificati","PeriodicalId":10883,"journal":{"name":"Cytometry Part B: Clinical Cytometry","volume":"110 1","pages":"7-10"},"PeriodicalIF":2.7,"publicationDate":"2026-01-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/cytob.70007","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146092413","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
{"title":"\"What are we measuring?\" reconsidering the biological identity and clinical interpretation of malaria-derived particles.","authors":"Zhihao Lei","doi":"10.1002/cytob.70011","DOIUrl":"https://doi.org/10.1002/cytob.70011","url":null,"abstract":"","PeriodicalId":10883,"journal":{"name":"Cytometry Part B: Clinical Cytometry","volume":" ","pages":""},"PeriodicalIF":2.7,"publicationDate":"2026-01-24","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146040581","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Alice Yue, Ryan R Brinkman, Veronica Nash, Fabian Junker, Goce Bogdanoski, Anagha Divekar, Aaron Tyznik, Josef Spidlen, Wolfgang Kern, Jordi Petriz, Kaska Wloka, Kamila Czechowska
{"title":"Response to \"Bridging the implementation gap in AI-assisted flow cytometry\".","authors":"Alice Yue, Ryan R Brinkman, Veronica Nash, Fabian Junker, Goce Bogdanoski, Anagha Divekar, Aaron Tyznik, Josef Spidlen, Wolfgang Kern, Jordi Petriz, Kaska Wloka, Kamila Czechowska","doi":"10.1002/cytob.70009","DOIUrl":"https://doi.org/10.1002/cytob.70009","url":null,"abstract":"","PeriodicalId":10883,"journal":{"name":"Cytometry Part B: Clinical Cytometry","volume":" ","pages":""},"PeriodicalIF":2.7,"publicationDate":"2026-01-21","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146009141","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Quantifying malarial parasite density is crucial for diagnosis and treatment in endemic areas. While Malaria-derived particles (MDPs) have been linked to malaria pathology, a direct quantification method for routine laboratory use remains unestablished. To address this, our study optimized a flow cytometry approach to enumerate MDPs per microliter of blood. Specimens were incubated with propidium iodide and red blood cell (RBC) lysis solution. The number of MDPs was quantified using a CytoFlex flow cytometer, size-standard beads, and counting beads. Electron microscopy was used to study the ultrastructures of the malarial parasites in the lysed RBC specimens. A significant increase in MDP levels was detected in blood samples from P. falciparum and P. vivax infections, but fewer than 1 particle/μL of MDPs were detected in the controls. The number of MDPs correlated with the percentage of infected red blood cells (iRBCs) obtained by manual counting (R2 = 0.94). The dilution assay demonstrated a strong correlation between the measured and expected values of the MDPs. An electron microscopic study demonstrated that different stages of malarial parasites exist in lysed RBCs in the form of membrane-bound spherical cells. A positive association was established between parasite density and MDPs across both P. falciparum (R2 = 0.94) and P. vivax (R2 = 0.91) infections. We demonstrated the potential use of flow cytometry for determining the MDP concentration. The developed approach is reliable and straightforward for the diagnosis and treatment of patients with malarial parasite infection in routine laboratory settings.
{"title":"A novel flow cytometry approach to quantify malaria-derived particles.","authors":"Attakorn Palasuwan, Kritsamon Sophondilok, Sumate Ampawong, Suttipat Srisutham, Egarit Noulsri, Duangdao Palasuwan","doi":"10.1002/cytob.70006","DOIUrl":"https://doi.org/10.1002/cytob.70006","url":null,"abstract":"<p><p>Quantifying malarial parasite density is crucial for diagnosis and treatment in endemic areas. While Malaria-derived particles (MDPs) have been linked to malaria pathology, a direct quantification method for routine laboratory use remains unestablished. To address this, our study optimized a flow cytometry approach to enumerate MDPs per microliter of blood. Specimens were incubated with propidium iodide and red blood cell (RBC) lysis solution. The number of MDPs was quantified using a CytoFlex flow cytometer, size-standard beads, and counting beads. Electron microscopy was used to study the ultrastructures of the malarial parasites in the lysed RBC specimens. A significant increase in MDP levels was detected in blood samples from P. falciparum and P. vivax infections, but fewer than 1 particle/μL of MDPs were detected in the controls. The number of MDPs correlated with the percentage of infected red blood cells (iRBCs) obtained by manual counting (R<sup>2</sup> = 0.94). The dilution assay demonstrated a strong correlation between the measured and expected values of the MDPs. An electron microscopic study demonstrated that different stages of malarial parasites exist in lysed RBCs in the form of membrane-bound spherical cells. A positive association was established between parasite density and MDPs across both P. falciparum (R<sup>2</sup> = 0.94) and P. vivax (R<sup>2</sup> = 0.91) infections. We demonstrated the potential use of flow cytometry for determining the MDP concentration. The developed approach is reliable and straightforward for the diagnosis and treatment of patients with malarial parasite infection in routine laboratory settings.</p>","PeriodicalId":10883,"journal":{"name":"Cytometry Part B: Clinical Cytometry","volume":" ","pages":""},"PeriodicalIF":2.7,"publicationDate":"2026-01-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145910885","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2026-01-01Epub Date: 2025-11-17DOI: 10.1002/cyto.b.22267
Devasis Panda, Narender Tejwani, Sabina Langer
{"title":"Coexistent dual CD4+/CD8- and CD4-/CD8+ T-cell large granular lymphocytic leukemia in a young adult male: An exceedingly rare finding.","authors":"Devasis Panda, Narender Tejwani, Sabina Langer","doi":"10.1002/cyto.b.22267","DOIUrl":"10.1002/cyto.b.22267","url":null,"abstract":"","PeriodicalId":10883,"journal":{"name":"Cytometry Part B: Clinical Cytometry","volume":" ","pages":"57-59"},"PeriodicalIF":2.7,"publicationDate":"2026-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145539464","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Vandana Panakkal, Raniah Al Amri, Stacey Mamatas, Sara A Monaghan, Ahmad Al-Attar
{"title":"Authors' Response to Letter from Khanolkar and Ahmed concerning our article \"An unusual pattern observed upon the addition of CD79b to a flow cytometry B-cell lymphoma panel\" (Panakkal et al., 2025). Cytometry Part B: Clinical Cytometry. DOI: 10.1002/cyto.b.22246. PMID: 40657818.","authors":"Vandana Panakkal, Raniah Al Amri, Stacey Mamatas, Sara A Monaghan, Ahmad Al-Attar","doi":"10.1002/cytob.70005","DOIUrl":"https://doi.org/10.1002/cytob.70005","url":null,"abstract":"","PeriodicalId":10883,"journal":{"name":"Cytometry Part B: Clinical Cytometry","volume":" ","pages":""},"PeriodicalIF":2.7,"publicationDate":"2025-12-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145827158","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Antibody titration is essential for optimizing platelet flow cytometry, a technique widely used to evaluate platelet phenotype, activation status, and function. This manuscript outlines practical approaches for platelet antibody titration in whole blood, with tailored strategies for constitutive and inducible markers. It emphasizes the use of appropriate controls, consideration of marker coexpression, and selection of subsaturating antibody concentrations to maximize signal resolution while minimizing background and artifactual activation. Quantitative metrics such as the stain index and separation index are introduced as tools for evaluating staining performance. The discussion also addresses key technical variables, including combinatorial titration, spillover spreading, lot variability, and antibody-induced activation. Titration under final assay conditions is recommended to ensure reproducibility and biological relevance. These strategies provide a foundation for developing robust, high-resolution platelet assays that support both research and clinical applications, particularly as flow cytometry evolves toward greater automation and standardization.
{"title":"Signal without noise: Practical antibody titration for platelet flow cytometry.","authors":"Benjamin E J Spurgeon","doi":"10.1002/cytob.70004","DOIUrl":"https://doi.org/10.1002/cytob.70004","url":null,"abstract":"<p><p>Antibody titration is essential for optimizing platelet flow cytometry, a technique widely used to evaluate platelet phenotype, activation status, and function. This manuscript outlines practical approaches for platelet antibody titration in whole blood, with tailored strategies for constitutive and inducible markers. It emphasizes the use of appropriate controls, consideration of marker coexpression, and selection of subsaturating antibody concentrations to maximize signal resolution while minimizing background and artifactual activation. Quantitative metrics such as the stain index and separation index are introduced as tools for evaluating staining performance. The discussion also addresses key technical variables, including combinatorial titration, spillover spreading, lot variability, and antibody-induced activation. Titration under final assay conditions is recommended to ensure reproducibility and biological relevance. These strategies provide a foundation for developing robust, high-resolution platelet assays that support both research and clinical applications, particularly as flow cytometry evolves toward greater automation and standardization.</p>","PeriodicalId":10883,"journal":{"name":"Cytometry Part B: Clinical Cytometry","volume":" ","pages":""},"PeriodicalIF":2.7,"publicationDate":"2025-12-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145780612","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Wei J Wang, Sa A Wang, Hong Fang, Qing Wei, Jeffrey L Jorgensen, Shimin Hu, Jie Xu, Shaoying Li, Guilin Tang, Zhenya Tang, L Jeffrey Medeiros, Wei Wang
{"title":"Challenges and approaches in the diagnosis and differential diagnosis of atypical CLL: A response to 'Defining atypical CLL for reproducible diagnosis: Implications of the work by Wang et al.'","authors":"Wei J Wang, Sa A Wang, Hong Fang, Qing Wei, Jeffrey L Jorgensen, Shimin Hu, Jie Xu, Shaoying Li, Guilin Tang, Zhenya Tang, L Jeffrey Medeiros, Wei Wang","doi":"10.1002/cytob.70003","DOIUrl":"https://doi.org/10.1002/cytob.70003","url":null,"abstract":"","PeriodicalId":10883,"journal":{"name":"Cytometry Part B: Clinical Cytometry","volume":" ","pages":""},"PeriodicalIF":2.7,"publicationDate":"2025-12-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145773768","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
扫码关注我们
求助内容：
应助结果提醒方式：
京公网安备 11010802042870号