2026 Meeting at Wiley

​​The MetroFlow NY/NJ Flow Cytometry Users Group invites you to attend our annual meeting on October 23rd, 2026.

We are planning a full day of scientific presentations and are excited to be hosted by Wiley Center at 111 River St, Hoboken, NJ 07030.

• Talks will run from 9am-5pm, with ample time over breaks to meet with our corporate sponsors.

  
  

• The meeting will conclude with a wine & cheese reception with excellent opportunities for networking.

8:00 AM - 8:50 AM Registration & Breakfast

8:50 AM - 9:00 AM Introduction

Thomas Miller, President of Metroflow, Director of Scientific Operations, Jill Roberts Institute for Research in Inflammatory Bowel Disease and Gale and Ira Drukier Institute for Children’s Health at Weill Cornell Medicine

9:00 AM - 9:30 AM "From Discovery to Impact: How Wiley Brings Trusted Research to Life"

Steven Ottogalli, Wiley, Director, Partner Publishing & Kate Lavallee-Sharma,Wiley, Senior Publisher, Partner

9:30 AM - 10:00 AM "From Scientist to Editor: A Journey Behind the Scenes"

Yolanda D Mahnke, PhD, Founder, FlowKnowHow LLC

10:00 AM - 10:30 AM Break

10:30 AM - 11:00 AM "Reticulocytes as Unexpected Regulators of Humoral Immunity"

Krystalyn E. Hudson, PhD, Columbia University

11:00 AM - 11:30 AM "Human blood stem cells remember stress: functional heterogeneity decoded by flow cytometry”

Stephanie Xie, PhD, Scientist; Assistant Professor, UHN, Princess Margaret Cancer Centre, University of Toronto

11:30 AM - 12:30 PM Panel

12:30 PM - 2:00 PM Lunch

2:00 PM - 2:30 PM "The Make Your Own Flow Cytometer: Continued evolution of a unique educational resource"

William Telford, PhD, Senior Associate Scientist, National Cancer Institute

2:30 PM - 3:00 PM “Towards ‘No Cell Left Behind’: open-source toolsets to leverage Spectral Flow Cytometry’s full biological discovery potential”

David Rach, PhD, Post-doctoral Research Fellow, University of Maryland - Baltimore, Greenebaum Comprehensive Cancer Center - Flow Cytometry Shared Resource

3:00 PM - 3:30 PM "Research Flow Cytometry Core at Regeneron”

Ben Daniel PhD, Sr. Director & Jason Walker, Sr. Artisanal Flow Cytometrist, Regeneron

3:30 PM - 4:00 PM Break

4:00 PM - 4:30 PM “Practical Challenges in Flow Cytometric Monitoring of CAR-T Clinical Trials”

Nithi Selliah, Global Director, Flow cytometry, Cerba Research

4:30 PM - 5:00 PM “A validated 29-color Treg-centric spectral flow cytometry assay with cross-platform algorithm-assisted analysis for autoimmune studies”

Calin O. Marian, Principal Scientist, Translational Molecular Biomarkers, Merck Research Laboratories, Merck & Co

5:00 PM - 5:10 PM Closing Remarks

Thomas Miller, President of Metroflow, Director of Scientific Operations, Jill Roberts Institute for Research in Inflammatory Bowel Disease and Gale and Ira Drukier Institute for Children’s Health at Weill Cornell Medicine

5:10 PM - 7:00 PM Wine & Cheese

Abstracts

Steven Ottogalli and Kate Lavallee-Sharma

An overview of Wiley's history and organization, how we partner with esteemed societies to publish their journals and advance knowledge, and an overview of the publishing process and strategy.

Yolanda Mahnke

Every academic scientist is involved in the world of scientific publishing to some degree – if nothing else, as an author. My own journey has taken me from being a scientist and author to my current role as Senior Associate Editor of Cytometry Part A, ISAC’s journal, published by Wiley.

Over the years, my engagement with scientific publishing has expanded well beyond manuscript authorship and peer review. My interactions with Wiley have grown both in scope and depth as we work to strengthen the partnership between Cytometry Part A and Wiley, and, more broadly, the connection between the journal, ISAC, and its annual CYTO conference.

In this presentation, I will share some of the projects and initiatives I have been involved in along the way, offering a behind-the-scenes look at the different facets of scientific publishing and how my role as a scientist has evolved through these experiences.

Krystalyn E. Hudson

Red blood cell (RBC) transfusion is one of the most common therapeutic procedures performed worldwide and remains a cornerstone of care for patients with sickle cell disease (SCD). Although transfusions can be lifesaving, some recipients develop alloantibodies against donor RBC antigens, creating substantial challenges for future transfusion support and contributing to adverse outcomes in transfusion, pregnancy, and transplantation. Despite the clinical significance of RBC alloimmunization, the factors that govern why some antibody responses develop and persist remain poorly understood.

Recent work from our laboratory has identified reticulocytes, immature RBCs that are markedly elevated in SCD and other hemolytic disorders, as unexpected regulators of humoral immunity. This presentation will discuss evidence demonstrating that reticulocytes promote RBC alloantibody responses and will highlight how flow cytometric approaches have enabled the characterization of erythroid-immune cell interactions. Findings from flow cytometry, imaging flow cytometry, confocal microscopy, and intravital imaging have revealed previously unappreciated interactions between reticulocytes and immune cells, providing new insights into how stress erythropoiesis influences antibody responses. Collectively, these findings challenge the traditional view of reticulocytes as passive erythroid precursors and establish a new role for stress erythropoiesis in the regulation of adaptive immunity.

Stephanie Z. Xie

Hematopoietic stem cells (HSC) have enormous regeneration capacity (~10E11 cells daily in the human). Adult humans are estimated to have 50,000 to 200,000 HSC contributing to hematopoiesis at any one time. Understanding the functional diversity of human HSC across a lifetime is crucial for promoting healthy aging, advancing medicine, and improving therapeutic strategies for hematological disorders. Age-related increase in inflammation is well-recognized to be a major risk factor for many diseases including cancer, heart disease and diabetes. Aging also strongly increases the risk for HSC to acquire mutations, which results in expansion of mutated blood cells in the blood and in various tissues in the body, a pre-leukemia process called clonal hematopoiesis. Flow cytometry has long been a cornerstone for interrogating HSC heterogeneity. We set out to understand if HSC inflammation responses underpin aging-associated disease outcomes by developing inflammatory recovery xenograft models and isolating HSC for downstream single cell molecular profiling. We identified inflammatory memory in a subset of human hematopoietic stem cells (HSC-iM), a heritable epigenetic and transcriptional state characterized by quiescence and inflammation signatures. HSC-iM links environmental inflammatory exposures to stem cell fate and diverse human conditions, including infection, aging, clonal hematopoiesis (CH), and mortality risk. We will discuss the workflow, key technical considerations, data integration strategies, and insights gained into the mechanistic regulation of human HSC function after inflammatory challenge. This platform provides a powerful framework for dissecting stem cell biology at unprecedented resolution and uncovering novel biomarkers and therapeutic targets in normal and malignant human hematopoiesis.

William Telford

As flow cytometers become more automated and complex, it becomes more challenging to teach basic instrument operation principles in an effective way.  The student-built flow cytometer as an instructional tool has therefore remained a critical (and enjoyable) component of flow cytometry education since 1990 when John Martin, Mark Wilder and James Jett of Los Alamos National Laboratory (LANL) introduced the first Build Your Own Flow Cytometer lab module (aka the Build Lab) at the Annual Course in Flow Cytometry in 1990.  Workshop students built a functioning flow cytometer from individual structural, fluidic, optical and electronic parts, providing a unique (and fun) hands-on experience that demonstrate how a flow cytometer functions “under the hood”.  After 36 years the continuously updated Build Lab remains the most popular module in the program, most recently presented at the 49th Annual Course in Flow Cytometry in San Diego, with no end in sight. In 2013 we introduced the Make Your Own Flow (MYO) Cytometer laboratory, a smaller system that could be easily transported abroad as checked luggage. For over 12 years this system has visited over 35 cytometry workshops in twenty countries, training hundreds of students in the “nuts and bolts” of cytometer theory and operation.   A major redesign in 2022 resulted in a more robust, compact and reliable design.  It also realigned the system with one of its original philosophies: To create a teaching tool that others could build for their own institutions and programs, either by reproducing the original system or by inspiring others to build their own designs.   Almost all structural elements are now built by additive manufacturing using inexpensive consumer-level 3-D printers.  This results in better (and more original) custom design, lighter weight, more reliable operation and lower cost.The system continues to see frequent use in international workshops, most recently in Argentina, Ghana, Italy and Chile, and in ISAC programs like CYTO Youth.  Improvements to its design and expanded usage in cytometry education programs will help us keep hands-on instrument building as an important component of cytometry education.

David Rach

Spectral Flow Cytometry (SFC) with its increased analytical breadth and width, has been a game changer for researchers working with limited biospecimen, enabling greater profiling of immune systems, and responses to infection and immunization. While community adoption of the technology has been rapid, the capacity to analyze the resulting high-dimensional datasets has not similarly scaled.

While analysis of a single cell population remains essentially unchanged, to fully explore the acquired datasets requires the use of unsupervised and semi-supervised algorithmic approaches. Existing methods, many originally intended for use with mass cytometry and single-cell RNA seq data, often struggle with SFC datasets due to increased number of events, and batch effects arising from the uncertainty inherent in the unmixing process. Consequently, the biological discovery potential of the technology remains under-leveraged for most datasets, with existing analysis just skimming the surface.

To address these technology specific challenges, we have been developing open-source toolsets in R and Rust. These have allowed us to screen unmixing controls for tandem degradation and additional autofluorescence, as well as detect failures in instrument quality control. We highlight some of our on-going work in the context of a semi-supervised analysis of a rare clinical cohort of HIV-exposed uninfected (HEU) neonates, additionally leveraging existing tool-sets from the R/Bioconductor ecosystem to provide a more replicable framework in the analysis of multiple SFC panels.

All the while, we contemplate the broader question: “What good is any tool, if the vast majority of the community is unable to use it?”

Nithianandan Selliah

Chimeric Antigen Receptor T-cell (CAR-T) therapy has transformed the treatment landscape for hematologic malignancies and is rapidly expanding into new therapeutic indications. As both autologous and allogeneic CAR-T products advance through clinical development, flow cytometry remains a critical tool for monitoring cell therapy kinetics, persistence, phenotype, and immune reconstitution.

While autologous CAR-T monitoring is relatively straightforward, the emergence of allogeneic "off-the-shelf" CAR-T products has introduced new analytical challenges. Traditional flow cytometric approaches often require modification because allogeneic products may lack conventional T-cell markers such as CD3 following gene editing, necessitating alternative gating strategies and CAR-specific detection methods.

This presentation will highlight practical considerations and real-world challenges encountered during flow cytometric monitoring of CAR-T clinical trials. Topics will include strategies for identifying and enumerating CAR-T cells, differences between autologous and allogeneic monitoring approaches, selection of CAR detection reagents, and interpretation of complex immunophenotyping data. Special emphasis will be placed on challenges associated with TBNK analysis in allogeneic CAR-T studies, where CAR-T cells within the CD3-negative compartment can complicate accurate identification of NK-cell populations.

Calin O. Marian

Introduction

Regulatory T cells (Tregs) maintain immune tolerance and are therapeutic targets in autoimmunity. High-dimensional spectral flow cytometry allows detailed Treg subset analysis, improving understanding of Treg biology for enhanced therapies. We developed and fit for purpose validated a 29 color Treg centric assay for frozen PBMCs and applied cross platform, algorithm assisted analysis (Astrolabe Diagnostics, Ozette Technologies, TerraFlow) in samples from systemic lupus erythematosus (SLE), ulcerative colitis (UC), and atopic dermatitis (AD) to standardize Treg subset monitoring in autoimmune cohorts.

Methods

A 29‑color T‑cell/Treg profiling panel was validated on a Cytek Aurora spectral cytometer and optimized for frozen PBMCs. It integrates viability and lineage (e.g., Live/Dead, CD3, CD4, CD8), differentiation/trafficking (e.g., CD45RA/RO, CCR7), activation/co‑stimulation (e.g., ICOS, 4‑1BB, GITR), inhibitory checkpoints (e.g., PD‑1, CTLA‑4, LAG‑3, TIGIT), regulatory/Treg markers (e.g., FoxP3, CD25, CD127, HELIOS, GARP/LAP), and proliferation (Ki‑67) to comprehensively profile conventional T cells and Tregs, and includes NK cell coverage (e.g., CD56). A two tube design included an FMO control tube for positivity thresholds and a full stain tube for reportables. Standardized workflows encompassed viability staining, surface staining, fixation/permeabilization, intracellular staining, and spectral unmixing with FMO guided thresholds. The assay was validated using in-house stimulated frozen PBMC QC material (three replicates across two runs; ≤20% CV intra-run and overall), intra-assay sample precision in frozen PBMCs from three NHV and three SLE donors (three replicates within one run; ≤20% CV for overall mean and majority of samples, relaxed to ≤30% CV for analytes with <1% mean), and an LLOQ for CD4+CD8-CD25+CD127-FoxP3+ (%CD4) via EXP:FMO admixtures (1:2–1:16; triplicates; ≤30% CV and ±30% relative error). Additional disease PBMCs (three UC, three AD) were processed for information only. The unmixed data were imported into OMIQ and pre-processed using PeacoQC. Tregs were pre-gated as CD25+ FoxP3+ CD127low within CD45+CD3+CD4+CD8− live cells. The pre-gated Tregs were exported and analyzed using the algorithm-assisted platforms provided by Astrolabe Diagnostics, Ozette Technologies, and TerraFlow under recommended settings. Cross-platform outputs were compared qualitatively for biological alignment; platform differences were evaluated in terms of gating strategy, marker inclusion/pruning, and label resolution.

Results

QC precision met acceptance across sentinel analytes (overall %CVs ~7–20%). Intra-assay sample precision was acceptable for most analytes, with low %CVs for abundant subsets and higher variability confined to low-frequency populations (<1%). Inter-assay context indicated 101/121 parameters met precision acceptance across runs. The LLOQ for CD4+CD8-CD25+CD127-FoxP3+ (%CD4) was established at 1.54% with acceptable precision and accuracy (linearity R2 ≈ 0.96). Most failures reflected very low event counts or borderline positivity for rare subsets (exploratory by design). Despite non-identical phenotype strings, all three platforms converged on memory Treg architecture—CD45RO+ CD45RA− CD95+ backbone—and a consistent CD39− vs CD39+ split. Baseline/quiescent memory (CD39−, typically CD31−/CD38−) appeared across platforms; suppressive memory (CD39+) was robust and often CD38low. Functional activation states (GARP/LAP+, Helios+, HLA-DR+) were explicit in Ozette and TerraFlow and mapped via the memory/CD39 scaffold in Astrolabe. Partial two-way strict and three-way relaxed similarity between platforms was observed for CD38+ recent activation and naive/RTE splits, reflecting differences in gate strategies (global vs per-sample vs predefined Boolean), marker inclusion/pruning (e.g., CD31/CD38), and labeling resolution.

Conclusions

A validated 29 color spectral assay for cryopreserved PBMCs, paired with cross platform algorithm assisted analysis, enables reproducible, high dimensional Treg profiling with platform agnostic biological interpretation. Concordant recovery of memory architecture, CD39−/CD39+ divergence, and activation states supports utility for pharmacodynamic monitoring and exploratory biomarker discovery in autoimmune studies. Limitations include small cohort size and qualitative cross platform comparisons; larger studies and harmonized labeling ontologies are planned.