The genesis and evolution of bead-based multiplexing.

Multiplexed analysis has the advantage of allowing for simultaneous detection of multiple analytes in a single reaction vessel which reduces time, labor, and cost as compared to single-reaction-based detection methods. Microsphere-based suspension array technologies, such as the Luminex® xMAP® system, offer high-throughput detection of both protein and nucleic acid targets in multiple assay chemistries. After Luminex's founding in 1995, it quickly became the leader in bead-based multiplexing solutions. Today, xMAP Technology is the most widely adopted bead-based multiplexing platform with over 35,000 peer-reviewed publications, an installed base of approximately 15,500 instruments, and over 70 Luminex Partners offering more than 1300 research use kits as well as custom assay solutions. Because of the open architecture of the xMAP platform it has been implemented in a variety of applications that range from transplant medicine, biomarker discovery and validation, pathogen detection, drug discovery, vaccine development, personalized medicine, neurodegeneration, and cancer research.

A conversation with Leonard and Leonore Herzenberg.

Leonard and Leonore Herzenberg have left an indelible mark on the fields of immunology and cell biology, both in research and clinical aspects. They are perhaps best known for developing the technologies of fluorescence flow cytometry and hybridomas. Over six decades, they made a number of important and fundamental discoveries in lymphocyte biology by applying these technologies. During this era, they immersed themselves in the sociopolitical environment, interjecting scientific rationale into public discourse about McCarthyism, nuclear fallout, war, genetics, and other politically charged topics. Their unique philosophy has shaped their lives, their science, and ultimately the scientific community. In this Conversation, we explore some of these driving forces and the impact on the laboratory.

Flow Cytometry: The Glass Is Half Full.

Accompanied by a historical perspective of the field of cytometry, this introductory chapter provides a broad view of what flow cytometry can do; hence, the glass is half full.

Wallace H. Coulter: decades of invention and discovery.

Only a few inventors can be said to have made as great an impact on mankind as Wallace Coulter. His inquisitive mind and ability to see well beyond what existed served him well for 40 years of inventing. So many of the fundamental tools that exist today in the area of hematology were derived from or driven by Coulter's inventions that he could be called the most technological innovator in the field of modern hematology. In achieving these discoveries Wallace Coulter was clearly capable of visualizing future opportunities that few others recognized. His vision was combined with an uncanny ability to translate his ideas into products. He developed a large number of tools that shaped the fields of cytometry, image analysis, and industrial materials. His understanding of the future power of computation drove him to link these technologies in a unique way. In the end, Coulter shaped the technologies that ultimately drove hematology in a new direction, one that remains on a critical pathway linking technology innovation all the way to true translational impact. It was said of Henry Ford that "[h]e has no notion that wealth has made him great, and any one who is imprest merely by his wealth bores him. In his personal contacts he likes to dodge the subject. He would prefer to talk with a machinist about machinery, or with somebody who likes birds about birds. In these contacts, he asks no deference; and if he gets it, he suspects it is mere deference to wealth, and that ends his interest."(1) The same could be said of Wallace Coulter, who, like Ford, understood the concepts of mass production and customer service. Coulter had the ability to recognize the opportunity and fulfill the need for development of a blood-cell counter that could be placed in every pathology laboratory, and in so doing transformed a field from a qualitative to a quantitative environment. Every person who has ever entered a medical lab, hospital, or doctor's office has felt the impact of Coulter's discovery.

The Evolution of Single-Cell Analysis and Utility in Drug Development.

This review provides a brief history of the advances of cellular analysis tools focusing on instrumentation, detection probes, and data analysis tools. The interplay of technological advancement and a deeper understanding of cellular biology are emphasized. The relevance of this topic to drug development is that the evaluation of cellular biomarkers has become a critical component of the development strategy for novel immune therapies, cell therapies, gene therapies, antiviral therapies, and vaccines. Moreover, recent technological advances in single-cell analysis are providing more robust cellular measurements and thus accelerating the advancement of novel therapies.Graphical abstract.

Reimagining Flow Cytometric Cell Sorting.

In this review, a brief history of this unrivaled technology, flow cytometry, is provided, highlighting its past and present advances, with particular focus on "flow cell" technologies. Flow cytometry has truly revolutionized high-throughput single cell analysis, which has tremendous implications, from laboratory to the clinic. This technology embodies what is truly referred to as cross fertile research, merging the physical with the life sciences. This review introduces the recent notable advancements in flow cell technology. This advancement sees the complete removal of liquid sheath flow, which has advanced the technology with the possibility of both the reduction in its foot print, while also simplifying the flow cells explored in cytometry. Interestingly, the novel sheathless flow cell technology demonstrated herein has the flexibility for handling both heterogeneous cell populations and whole organisms, thus demonstrating a versatile flow cell technology for both flow cytometry and fluorescent-activated cell sorting.

Mass cytometry and type 1 diabetes research in the age of single-cell data science.

PURPOSE OF REVIEW: New single-cell tec. hnologies developed over the past decade have considerably reshaped the biomedical research landscape, and more recently have found their way into studies probing the pathogenesis of type 1 diabetes (T1D). In this context, the emergence of mass cytometry in 2009 revolutionized immunological research in two fundamental ways that also affect the T1D world: first, its ready embrace by the community and rapid dissemination across academic and private science centers alike established a new standard of analytical complexity for the high-dimensional proteomic stratification of single-cell populations; and second, the somewhat unexpected arrival of mass cytometry awoke the flow cytometry field from its seeming sleeping beauty stupor and precipitated substantial technological advances that by now approach a degree of analytical dimensionality comparable to mass cytometry. RECENT FINDINGS: Here, we summarize in detail how mass cytometry has thus far been harnessed for the pursuit of discovery studies in T1D science; we provide a succinct overview of other single-cell analysis platforms that already have been or soon will be integrated into various T1D investigations; and we briefly consider how effective adoption of these technologies requires an adjusted model for expense allocation, prioritization of experimental questions, division of labor, and recognition of scientific contributions. SUMMARY: The introduction of contemporary single-cell technologies in general, and of mass cytometry, in particular, provides important new opportunities for current and future T1D research; the necessary reconfiguration of research strategies to accommodate implementation of these technologies, however, may both broaden research endeavors by fostering genuine team science, and constrain their actual practice because of the need for considerable investments into infrastructure and technical expertise.