Improving care is a mission that drives us all. Translating new discoveries into better clinical outcomes takes dedication and new approaches to asking the most difficult questions. At Fluidigm, we believe that changing the course of how diseases are treated and ultimately cured requires a comprehensive understanding of complex cellular phenotypes and their interrelationships within the tissue microenvironment. To meet this great challenge, we developed the Hyperion™ Imaging System.

“The impact of this first-in-class technology stands to be especially profound in oncology, immunology and immunotherapy research. With more than 30 systems enabled globally, progress in applying this innovative technology to ask new questions across a number of disease research areas is truly impressive .”
—Chris Linthwaite, President and CEO, Fluidigm

The Hyperion Imaging System brings imaging capability together with proven CyTOF® technology to set a new standard in highly multiplexed protein detection. Surpassing the inherent limitations of fluorescence detection, this system enables researchers to simultaneously image 4 to 37 protein markers in tissues and tumors by Imaging Mass Cytometry™. Ideal for deep profiling of precious formalin-fixed, paraffin-embedded (FFPE) and fresh frozen tissue samples, the Hyperion Imaging System empowers researchers to uncover new biomarker correlations and cellular social networks in the context of the tissue microenvironment—all from a single slide.

As with any new research application, having access to optimized reagents, analytical tools and training is essential for success. We designed the Hyperion Imaging System workflow with this in mind. It is accompanied by a pathologist-verified Maxpar® imaging antibody portfolio, antibody labeling kits, a software suite for image acquisition and data analysis, and experienced Fluidigm service and support. With the ability to utilize up to 135 channels to detect additional parameters, the system is well-suited to meet researchers needs today and well into the future.

A step jump of this magnitude doesn’t happen every day in life sciences. The impact of this first-in-class technology stands to be especially profound in oncology, immunology and immunotherapy research. With more than 30 systems enabled globally, truly impressive progress is being made in applying this innovative technology to ask new questions across a number of disease research areas.

I invite you to join with us and the growing scientific community in powering this revolution in tissue imaging to create breakthroughs today that will enable the development of better diagnostics and more effective therapies for tomorrow.

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For Research Use Only. Not for use in diagnostic procedures.

The Hyperion™ Imaging System received first place in The Analytical Scientist Innovation Awards, or TASIAs. Judges named it the top innovation of 2017, commending the platform that “combines tissue imaging with cytometry, allowing correlation between biomarkers and cell interactions. This technology has the potential to be truly transformative.”

Editor Charlotte Barker of The Analytical Scientist said, “Now in year five, The Innovation Awards continue to shine a light on the instruments and technologies that are having a big impact across the analytical sciences. We had a record number of entries this year and the final 15 reflect the full spectrum of analytical advances, from ingenious software for managing impurity data, to transformative MS imaging technology.”

The Hyperion Imaging System has the potential to revolutionize disease research by providing unprecedented visualization of complex cellular phenotypes and their relationships in the context of cancer, immuno-oncology and a range of immune mediated diseases. The potential ability to develop better diagnostics and more effective therapies stands to change the future of health care.

Read the full article in The Analytical Scientist for more on the TASIAs. To learn more about Imaging Mass Cytometry™ and be informed about news and events, fill out the form below.

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For Research Use Only. Not for use in diagnostic procedures.

Watch the video to learn about the power of Imaging Mass Cytometry.

Hyperion Imaging System

The Hyperion™ Imaging System brings together imaging capability with proven CyTOF® technology to facilitate Imaging Mass Cytometry™ applications. This allows highly multiplexed immunohistochemistry enabling the simultaneous analysis of 4 to 37 protein markers. Deep interrogation of tissues or tumors at subcellular resolution can empower the identification of new prognostic and diagnostic biomarkers and provide key insights into disease progression, response to therapy or drug mechanisms in the future. Discover more about Imaging Mass Cytometry.

For news, updates and more information about Imaging Mass Cytometry and the Hyperion Imaging System, send us your contact details using the form below.

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For Research Use Only. Not for use in diagnostic procedures.

Mass cytometry is fast becoming an industry standard for deep cellular profiling in translational and clinical research, referenced by hundreds of peer-reviewed publications around the world.

Download this compendium of mass cytometry research applications to hear from six investigators who share how this transformative technology is allowing them to reveal new insights in health and disease. Within this compendium you’ll also find a brief history of mass cytometry, an overview of the technology and information on the high-dimensional data analysis options provided by Cytobank.

We hope you enjoy learning about the unique insights mass cytometry brought to the researchers below in the areas of hematology/oncology, tumor biology, autoimmune disease and basic immunology.

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Table of Contents for Insights Powered by Mass Cytometry

	Mass Cytometry for Discovery and Functional Profiling: An Introduction to the Technology Dmitry Bandura Fluidigm
	Unlocking the Potential of Mass Cytometry Sean Bendall Stanford University
	Discovering Normal to Better Understand Abnormal: Searching for an Improved ALL Diagnosis Dr. Kara Davis Stanford University
	Profiling T Cell Diversity in Solid Tumors with Mass Cytometry Dr. Kavita Dhodapkar Yale University School of Medicine
	Understanding Variation in the Human Immune System Dr. Petter Brodin Karolinska Institutet
	Tackling Dynamic Complexity to Understand Immunotherapy Responses in Multiple Sclerosis Sonia Gavasso University of Bergen
	Single-Cell Insights into AML Cancer Biology Dr. Zohar Sachs University of Minnesota
	Mass Cytometry Processing with Cytobank Hannah Polikowsky Cytobank

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Complete the form below to download the Mass Cytometry Compendium

For Research Use Only. Not for use in diagnostic procedures.

Tumor gene expression has proven effective in measuring immune response during cancer progression and therapeutic response. As emerging therapies reveal new biomarkers and expand the need for samples, the costs and labor required to complete this important work also rise. Cancer researchers now have a reliable, sensitive and cost-effective tool for identifying gene expression signatures from immune and cancer cells: the new Advanta™ Immuno-Oncology Gene Expression Assay, used with the Biomark™ HD system.

A powerful immuno-oncology focused assay

The Advanta Immuno-Oncology Gene Expression Assay was developed in collaboration with leading academic and biopharma researchers to provide the right balance of biomarker breadth, assay flexibility and workflow efficiency.

The panel consists of 170 genes, including 91 reported in Nature, where Roy Herbst et al. defined a gene set representing tumor and immune response. The panel includes markers for:

• Immune cell identification
• Immune and cancer cell function
• Immune regulation and cell fate
• Checkpoint therapy response

Optimized for FFPE and fresh frozen tumor samples, the Advanta Immuno-Oncology Gene Expression Assay uses TaqMan® chemistry to sensitively measure gene expression, with five reference genes serving as analysis controls. Combined with Fluidigm microfluidics technology, the assay uniquely offers significant workflow efficiencies over traditional gene expression profiling methods. Each reaction is miniaturized to nanoliter volume and controlled using precise automation to empower accurate and cost-effective qPCR analysis across a large dynamic range. Since assay introduction into the integrated fluidic circuit (IFC) is under user control, researchers also have the flexibility to add up to 17 new assays or exchange gene assays within the panel to achieve experimental goals—all without affecting the original panel content, protocol or workflow.

Immuno-Oncology Expression Assay

Panel A		Panel B
ARG1	CLEC4C	IL12A	PDCD1LG2 (PD-L2)		APOBEC3A	CXCR4	IFNA2	NKG7
BTLA	CSF2	IL13	PRF1		APOBEC3B	CYBB	IGHA1	NRAS
CCL2	CTLA4	IL17A	PTGER2		ARG2	DGAT2	IGHG1	NT5E
CCL22	CX3CL1	IL17F	PTGER4		CA4	EBI3	IGHM	PYGL
CCL28	CXCL10	IL1B	PTGS2		CCL18	ERBB2	JCHAIN (IGJ)	SLAMF7
CCR5	CXCL8	IL2	PTPRC		CCL21	FASLG	IGKC	SLAMF8
CCR7	CXCL9	IL2RA	RORC		CCL3	FCER1G	IGLJ3	STAT1
CD1C	CXCR3	IL4	SDHA		CCL4	FCRLA	IGSF6	STAT2
CD244	EOMES	IL6	SP2		CCL5	FYB	IL10RA	STAT3
CD27	EPCAM	IL7	TBX21		CD160	GATA3	IL12B	STAT5A
CD274 (PD-L1)	FOXP3	IL7R	TGFB1		CD19	GNLY	IL15	STAT5B
CD276	GZMA	ITGAM	TMEM55B		CD1D	GZMH	IL2RG	STAT6
CD28	GZMB	ITGAX	TNF		CD2	GZMK	IRF9	TLR7
CD3E	HAVCR2	ITGB2	TNFRSF14		CD22	HLA-DMB	ISG15	TLR8
CD4	HLA-A	KLRK1	TNFRSF4		CD37	HLA-DPB1	JAK2	TNFAIP8
CD40	HLA-B	LAG3	TNFRSF9		CD52	HLA-DQB1	KREMEN1	TNFRSF18
CD40LG	HLA-C	LGALS9	TNFSF4		CD53	HLA-DRB1	LAPTM5	TNFSF18
CD48	HMOX1	MAP4K1	TNFSF9		CD63	IFI27	LCK
CD69	ICAM1	MICA	VCAM1		CTSS	IFIT2	LRG1
CD70	ICOS	MICB	VEGFA
CD80	IDO1	MS4A1	VPS33B
CD86	IFNG	NCAM1	VTCN1
CD8A	IL10	PDCD1 (PD-1)			Housekeeping genes for panels A and B: B2M, ACTB, GUSB, TFRC

Table 1. The Advanta Immuno-Oncology Gene Expression Assay panels A and B. This two-panel set includes 91 target genes and 5 housekeeping genes on Panel A and 74 target genes and the same 5 housekeeping genes on Panel B. Panel B reserves 17 open assay inlets, so you have the added flexiblity to add new assays over time for customization. Each panel is available in packs of 2 or 10 (192 or 960 samples, respectively)

Sample case study

In a 2016 paper published by The Lancet for the POPLAR study, “Atezolizumab versus docetaxel for patients with previously treated non-small-cell lung cancer,” Louis Fehrenbacher et al. detailed how they “assessed efficacy and safety of atezolizumab versus docetaxel in previously treated NSCLC, analyzed by PD-L1 expression levels on tumor cells and tumor-infiltrating immune cells and in the intention-to-treat population.” The team showed that “patients with pre-existing immunity, defined by high T-effector–interferon-γ-associated gene expression, had improved overall survival with atezolizumab.”

Fehrenbacher’s group noted that atezolizumab “significantly improved survival compared with docetaxel in patients with previously treated NSCLC. Improvement correlated with PD-L1 immunohistochemistry expression on tumor ce/p>lls and tumor-infiltrating immune cells,” leading them to conclude: “PD-L1 expression is predictive for atezolizumab benefit. Atezolizumab was well tolerated, with a safety profile distinct from chemotherapy.”

The researchers performed the clinical trial using immunohistochemistry, and also ran a gene expression panel of 91 genes featured in the Herbst paper. These genes are included in the Advanta Immuno-Oncology Gene Expression Assay. The team subsequently used these results to develop a gene signature based on a subset of the panel, effectively predicting therapeutic response that provides data that could potentially be used for developing future diagnostics tests.

Achieve greater overall productivity

The Advanta Immuno-Oncology Gene Expression Assay is optimized to run on the industry-proven Biomark HD system. This automated system enables researchers to easily run up to 96 samples at once with limited hands-on time, while delivering trusted performance. Together with the capability to sensitively detect biomarkers across defined T cell subsets, immune regulation, immune cell fate, cytokines, chemokines and more with exceptional cost-efficiency, this new assay offers an ideal solution to meet the rigorous demands of translational research studies.

Advanta IO Gene Expression Assay ordering information

Product Name	Contents	Part Number
Advanta Immuno-Oncology Gene Expression Assay Panel A-2 IFCs	Reagents, assay, syringes, IFCs	101-6082
Advanta Immuno-Oncology Gene Expression Assay Panel B-2 IFCs	Reagents, assay, syringes, IFCs	101-6083
Advanta Immuno-Oncology Gene Expression Assay Panel A & B-4 IFCs	Reagents, assay, syringes, IFCs	101-6084

Contact us to learn more about the Biomark HD system and the Advanta Immuno-Oncology Gene Expression Assay.

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For Research Use Only. Not for use in diagnostic procedures.

Every year since 2012, the mass cytometry community has gathered to share its research findings and technical know-how in an atmosphere fostering support and collaboration. On June 9 of this year, the Museum of Science in Boston was the scene for the 6th Annual Mass Cytometry Summit, our largest ever with more than 200 attendees.

Watch a video of mass cytometry users sharing their thoughts on the Summit.

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“The mass cytometry community is very dynamic and people are really open to exchanging ideas and protocols. Initially, most mass cytometry information was coming from the US. Now you can find people interacting all across the globe.”
—Hervé Luche, R&D manager, Centre d'ImmunoPHEnomique, Marseille

Researchers presented talks on topics ranging from new mass cytometry applications to best practices and data analysis approaches. Highlights included Ruth Montgomery from Yale and Stephen Oh from Washington University presenting on their use of mass cytometry in translational studies; Adeeb Rahman from Icahn School of Medicine at Mount Sinai presenting his innovative approach for a mass cytometric surrogate of light scatter; and Vinko Tosevski from the University of Zurich sharing his facility’s highly effective workflow for Cytobank data analysis. An informative panel discussion sparked lively audience engagement on integrating mass cytometry services into both academic and pharmaceutical institutions.

Jonathan Irish, PhD, Vanderbilt University, led the panel discussion “Meeting the Challenge of Integrating Mass Cytometry at Your Institution” at the 6th Annual Mass Cytometry Summit. Panelists included, from left to right, Ruth Montgomery, PhD, Yale School of Medicine; Mike Lee, MS, University of California, San Francisco; Carina Torres, BS, Eli Lilly; Mike Leipold, PhD, Stanford University; John Daley, BS, Dana-Farber Cancer Institute.

Scientists from around the world participated, representing top-tier research institutions and major pharmaceutical companies. Attendees interacted and networked during breaks and meals on the banks of the Charles River at the Museum of Science outdoor pavilion.

“The Mass Cytometry Summit gives a unique way to interact with other researchers in a relaxed environment: It’s easy to meet new people and establish collaborations.”
—Denis Schapiro, PhD, University of Zurich

Many speakers graciously agreed to share their talks with the mass cytometry community. Watch their on-demand presentations.

Watch

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For Research Use Only. Not for use in diagnostic procedures.

Enabling multiparameter profiling of precious samples, mass cytometry has been instrumental in achieving new research breakthroughs in areas such as immunology, cancer immunotherapy, drug discovery, vaccine development and more around the world.

Watch the webinars below to learn how researchers are utilizing Helios™, a CyTOF system, and mass cytometry applications to further their research.

	Powering multi-omic translational studies with mass cytometry Brice Gaudilliere, MD, PhD  Assistant Professor, Department of Anesthesiology Stanford University School of Medicine Nima Aghaeepour, PhD Assistant Professor Stanford University School of Medicine Watch
	Finding Pathologic T Cells in Rheumatoid Arthritis by Mass Cytometry Deepak Rao, MD, PhD  Rheumatologist Co-Director, Human Immunology Center Brigham and Women's Hospital Watch
	Querying Cytokine-Signaling Networks in Myeloproliferative Neoplasms Stephen Oh, MD, PhD  Assistant Professor of Medicine, Division of Hematology  Washington University School of Medicine  Watch
	Enhancing Checkpoint Blockade in Lymphoma with In Situ Vaccination Joshua Brody, MD  Assistant Professor, Hematology and Medical Oncology Director, Lymphoma Immunotherapy Program Icahn School of Medicine at Mount Sinai Watch
	Decoding Tumors with Mass Cytometry Jonathan Irish, PhD Assistant Professor of Cancer Biology, Assistant Professor of Pathology, Microbiology and Immunology Vanderbilt University Watch
	Defining the Phenotypic and Functional Immune Microenvironment in AML Evan Lind, PhD Assistant Professor, Department of Molecular Microbiology and Immunology and Department of Cell, Development and Cancer Biology Oregon Health & Science University Watch
	Towards an Assay of Global Immune Competence Holden Maecker, PhD Associate Professor (Research) of Microbiology and Director, Human Immune Monitoring Center Stanford University Watch
	Signatures of Protective NK Cell Responses Catherine Blish, MD, PhD Assistant Professor, Department of Medicine and Immunology, Stanford University School of Medicine Assistant Director of the Stanford Medical Scientist Training Program Watch

Fluidigm

For Research Use Only. Not for use in diagnostic procedures.

The C1™ system provides the widest range of automated single-cell applications in the industry, from gene expression profiling to genomic and epigenetic analysis. With more than 150 peer-reviewed publications, the C1 system has empowered significant research breakthroughs in the understanding of cell diversity and function in cancer, neurology, immunology and cardiovascular and stem cell biology.

The key to successful single-cell studies is preparing high-quality suspensions with maximum cell viability and minimum cell stress and debris. Whether you are working with cultured cells or with samples freshly isolated from tissue, cell quality is a critical factor for ensuring optimal results in single-cell datasets.

Now you can load enriched cell populations obtained by flow cytometric cell sorting directly into the C1 IFC (integrated fluidic circuit) to obtain rare subpopulations of cells with high purity and viability.

Benefits of this new application include:

•	A streamlined workflow that’s gentler and reduces cell losses as compared to sorting into a microfuge tube and then transferring to C1.
•	Reduced sample processing time.
•	Improved capture efficiency for neurons and immune cells.

To learn more, download the application note Cell Sorting Directly to the C1 IFC and the associated C1 cell sorting workbook today.

For Research Use Only. Not for use in diagnostic procedures.

The promising field of immuno-oncology presents many unanswered questions for scientists in biopharma, cancer research, flow and mass cytometry core facilities and immunology and immuno-oncology academic institutions: How do cancer cells evade the immune system? Which cells and pathways are the best molecular targets? What is the mechanism of action for potential pharmaceuticals? How can we increase tumor immunogenicity? Which biomarkers could predict response? Which drugs offer the best treatment outcomes, and in what order of use?

Approaches based on immunotherapy are providing unprecedented benefits for cancer patients—most importantly, durable responses and complete remission at rates that far surpass conventional chemotherapy. The defining feature of immunotherapy is that T cells and other cell types in the immune system mediate the anticancer response. But immune system cells also promote adverse reactions in some immunotherapy patients. Monitoring T cells and other immune cells in these patients is important for drug development, understanding mechanisms of action and determining biomarkers that correlate with both response and adverse events.

“Enabling the immune system to recognize and eliminate tumors has tremendous potential to change the way cancer is treated and eventually cured. Using the unique power of mass cytometry, the immuno-oncology research community can identify new biomarkers and optimize the development of new cancer immunotherapies to fully realize the potential of this transformative approach to cancer care.”
—Chris Linthwaite, President and CEO, Fluidigm

The Maxpar® Human T Cell Immuno-Oncology EX Panel Kit—together with the Human T Cell Phenotyping Panel Kits—enables comprehensive identification of all major T cell subsets for determining activation, homing status and checkpoint molecule expression. Designed for use with mass cytometry, with capabilities for measuring more than 40 parameters per cell with little or no signal overlap, these panels reveal highly valuable information from even the most limited biological samples.

Modular configuration makes it easy to combine T Cell Phenotyping Panel Kits for flexible or complete coverage. Individual antibodies offer further customization for specific research needs. With access to a growing catalog of more than 600 commercially available preconjugated antibodies and custom conjugation options, the Maxpar panel system can be adapted to meet changing needs in cancer immunotherapy research.

A new industry standard

“The dynamic nature and inherent complexity of the antitumor immune response requires new approaches to deeply profile cell populations at single-cell resolution,” said Fluidigm President and CEO Chris Linthwaite. “This new set of multiparameter panels will empower more researchers with high-dimensional tools to interrogate the tumor immune response and the tumor microenvironment, maximizing the information obtained from each precious sample.”

“Enabling the immune system to recognize and eliminate tumors has tremendous potential to change the way cancer is treated and eventually cured,” Linthwaite added. “Using the unique power of mass cytometry, the immuno-oncology research community can identify new biomarkers and optimize the development of new cancer immunotherapies to fully realize the potential of this transformative approach to cancer care.”

Mass cytometry is fast becoming an industry standard for deep cellular profiling in translational and clinical research. Referenced by hundreds of peer reviewed publications, mass cytometry expands beyond the inherent technical limitations of flow cytometry. With minimal spectral overlap, the technology enables researchers to interrogate more than 40 parameters simultaneously from millions of cells across both intracellular and extracellular protein markers.

Sample case study

As an example, The Journal of Immunology published a paper by Rituparna Das et al., “Combination therapy with anti-CTLA-4 and anti-PD-1 leads to distinct immunologic changes in vivo,” demonstrating highly multiplexed phenotyping of circulating immune cells to characterize immune response to individual and combination therapies. The report detailed the team’s experimental approach in a 45-patient cancer study at Yale University and Memorial Sloan Kettering Cancer Center.

Das and her collaborators used mass cytometry to perform multiparameter immunophenotyping of human PBMCs from patients before and after they were administered anti-PD-1 and/or anti-CTLA-4 checkpoint blockade therapy. Their customized 20-marker panel included the 16-marker Maxpar Human T Cell Phenotyping Panel Kit. They reported that the population of Ki-67+ cells increasing after anti-CTLA-4 or combination checkpoint blockade had a phenotype of CD45RO+, CCR7−, CD27+, CD28+ and CD95+, “consistent with transitional memory T cells.” These same proliferating cells also upregulated MHC class II HLA-DR.

Such deep profiling and identification of changes in this unique cell population would have been difficult using traditional approaches. Our new T cell immuno-oncology panel sets offer even deeper profiling than those used in the study cited above. Combining Maxpar T cell and immuno-oncology panels creates an unprecedented tool for profiling T cells. These panel sets also illustrate the power of mass cytometry as the premier platform for immune profiling and monitoring applications in translational and clinical research.

Register for the 6th Annual Mass Cytometry Summit in Boston on June 9 to learn about recent insights made using mass cytometry and the Maxpar Human T Cell Immuno-Oncology EX Panel Kit.

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For Research Use Only. Not for use in diagnostic procedures.

As single-cell research expands around the globe, researchers are profiling greater numbers of cells to identify new cellular subtypes. Scientists can then explore the role and function of each cell population in health and disease states by characterizing their genotypic, transcriptional, epigenetic and proteomic phenotypes. The C1™ system supports more automated single-cell applications than any other platform, enabling greater understanding of cellular diversity and function across many areas of disease research.

The new C1 Single-Cell mRNA Seq HT IFC is designed to expand current understanding of cellular heterogeneity by empowering scientists studying cancer, stem cells and organismal development with better research capabilities, improved single-cell capture and high-efficiency workflow performance. Isolate up to 800 medium size (10–17 µm) single cells for mRNA sequencing analysis.

“The release of the new medium-cell HT IFC with higher single-cell capture performance is exciting. With this new offering on the C1 system, researchers have a powerful new tool to explore the diversity of cell populations at single-cell resolution, discover rare and novel cell types and characterize their role in life.”
—Chris Linthwaite, Fluidigm President and CEO

Improved reagents, better gene detection

Together with the C1 Single-Cell mRNA Seq HT Reagent Kit v2, the medium HT IFC (high throughput integrated fluidic circuit) detects thousands of genes per cell using 3′ end counting and delivers excellent sensitivity, with design elements that provide powerful control over experimental costs and data quality. New HT product pricing makes this advanced technology more accessible while enabling larger single-cell studies.

“The release of the new medium-cell HT IFC with higher single-cell capture performance is exciting,” said Fluidigm President and CEO Chris Linthwaite. “With this new offering on the C1 system, researchers have a powerful new tool to explore the diversity of cell populations at single-cell resolution, discover rare and novel cell types and characterize their role in life.”

Clear insights

The new medium-cell HT IFC empowers researchers to discover and characterize critical cell populations that remain virtually invisible in the absence of single-cell resolution. It’s a development that promises to reveal fresh scientific insights into the biological roles of rare cell types.

Learn more about the C1 system and the power of microfluidics with single-cell IFCs.

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For Research Use Only. Not for use in diagnostic procedures.