RESUMO
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.
Assuntos
Corantes Fluorescentes , Citometria de Fluxo/históriaRESUMO
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.
Assuntos
Desenvolvimento de Medicamentos/tendências , Citometria de Fluxo/tendências , Análise de Célula Única/tendências , Desenvolvimento de Medicamentos/história , Desenvolvimento de Medicamentos/métodos , Citometria de Fluxo/história , Citometria de Fluxo/métodos , História do Século XVI , História do Século XVII , História do Século XVIII , História do Século XIX , História do Século XX , História do Século XXI , Humanos , Microscopia/história , Microscopia/métodos , Microscopia/tendências , Análise de Célula Única/história , Análise de Célula Única/métodosRESUMO
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.
Assuntos
Pesquisa Biomédica/tendências , Ciência de Dados/tendências , Diabetes Mellitus Tipo 1/etiologia , Proteômica/métodos , Análise de Célula Única/tendências , Animais , Pesquisa Biomédica/história , Pesquisa Biomédica/métodos , Ciência de Dados/história , Ciência de Dados/métodos , Diabetes Mellitus Tipo 1/patologia , Citometria de Fluxo/história , Citometria de Fluxo/métodos , Citometria de Fluxo/tendências , História do Século XXI , Humanos , Espectrometria de Massas/história , Espectrometria de Massas/métodos , Espectrometria de Massas/tendências , Proteômica/história , Proteômica/tendências , Análise de Célula Única/história , Análise de Célula Única/métodosRESUMO
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.
Assuntos
Separação Celular/métodos , Citometria de Fluxo/métodos , Análise de Célula Única/métodos , Separação Celular/história , Separação Celular/instrumentação , Citometria de Fluxo/história , Citometria de Fluxo/instrumentação , Corantes Fluorescentes/química , História do Século XX , História do Século XXI , Humanos , Hidrodinâmica , Lasers , Fibras Ópticas , Reologia/métodos , Análise de Célula Única/história , Análise de Célula Única/instrumentaçãoAssuntos
Biologia Celular/organização & administração , Congressos como Assunto , Citometria de Fluxo , Citometria por Imagem , Invenções , Sociedades Científicas/organização & administração , Canadá , Biologia Celular/economia , Biologia Celular/história , Biologia Celular/tendências , Congressos como Assunto/história , Congressos como Assunto/organização & administração , Congressos como Assunto/tendências , Técnicas Citológicas/história , Técnicas Citológicas/métodos , Técnicas Citológicas/tendências , República Tcheca , Indústria Farmacêutica/organização & administração , Indústria Farmacêutica/tendências , Educação/história , Educação/organização & administração , Educação/tendências , Citometria de Fluxo/história , Citometria de Fluxo/métodos , Citometria de Fluxo/tendências , Obtenção de Fundos/organização & administração , Obtenção de Fundos/tendências , História do Século XXI , Humanos , Citometria por Imagem/história , Citometria por Imagem/métodos , Citometria por Imagem/tendências , Invenções/economia , Invenções/tendências , Técnicas Analíticas Microfluídicas/instrumentação , Técnicas Analíticas Microfluídicas/métodos , Técnicas Analíticas Microfluídicas/tendências , Empresa de Pequeno Porte/economia , Empresa de Pequeno Porte/métodos , Empresa de Pequeno Porte/organização & administração , Empresa de Pequeno Porte/tendências , Sociedades Científicas/economia , Sociedades Científicas/história , Sociedades Científicas/tendênciasRESUMO
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.
Assuntos
Ensaios de Triagem em Larga Escala/história , Microesferas , Biomarcadores/análise , Ensaios Enzimáticos/história , Ensaios Enzimáticos/instrumentação , Ensaios Enzimáticos/métodos , Ensaios Enzimáticos/tendências , Citometria de Fluxo/história , Citometria de Fluxo/instrumentação , Citometria de Fluxo/métodos , Citometria de Fluxo/tendências , Ensaios de Triagem em Larga Escala/instrumentação , Ensaios de Triagem em Larga Escala/métodos , Ensaios de Triagem em Larga Escala/tendências , História do Século XX , História do Século XXI , Humanos , Imunoensaio/história , Imunoensaio/instrumentação , Imunoensaio/métodos , Imunoensaio/tendências , Fenômenos Magnéticos , Hibridização de Ácido NucleicoRESUMO
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.
Assuntos
Citometria de Fluxo , Citometria de Fluxo/história , Citometria de Fluxo/métodos , Citometria de Fluxo/tendências , História do Século XX , História do Século XXI , HumanosRESUMO
This chapter describes advantages and limitations of imaging flow cytometry (IFC) based on Imagestream instrumentation using a hybrid approach of morphometric measurement and quantitation of multiparametric fluorescent intensities' distribution in cells and particles. Brief comparison is given of IFC with conventional flow cytometry and fluorescent microscopy. Some future directions of the IFC technology are described and discussed.
Assuntos
Citometria de Fluxo/métodos , Citometria por Imagem/métodos , Citometria de Fluxo/história , Citometria de Fluxo/instrumentação , História do Século XX , História do Século XXI , Citometria por Imagem/história , Citometria por Imagem/instrumentação , Microscopia de Fluorescência , SoftwareRESUMO
The onset of the AIDS pandemic in the early 1980s coincided with the convergence of technologies now collectively known as flow cytometry (FCM). Major advances in FCM led significantly toward our understanding of the pathogenicity of the disease, which in turn led to wider adoption of the technology, including using it effectively in a variety of diagnostics. CD4+ T lymphocyte population counts, along with human immunodeficiency virus (HIV) viral load, remain the gold standard in diagnosis and continue to play a major role in the monitoring of advanced retroviral therapies. Arguably, the spread of AIDS (acquired immunodeficiency syndrome), the HIV virus, and the toll of the virus on humanity have been considerably altered by the concurrent development of FCM, the details of which are presented herein.
Assuntos
Síndrome da Imunodeficiência Adquirida/diagnóstico , Síndrome da Imunodeficiência Adquirida/epidemiologia , Citometria de Fluxo/métodos , Pandemias , Contagem de Linfócito CD4 , Feminino , Citometria de Fluxo/história , Citometria de Fluxo/tendências , História do Século XX , Humanos , MasculinoAssuntos
Células Produtoras de Anticorpos/citologia , Baço/citologia , Animais , Células Produtoras de Anticorpos/imunologia , Células CHO , Cricetinae , Cricetulus , Citometria de Fluxo/história , Citometria de Fluxo/instrumentação , Citometria de Fluxo/métodos , História do Século XX , Camundongos , Ovinos , Baço/imunologiaAssuntos
Linfócitos B , Citometria de Fluxo , Baço , Células-Tronco , Linfócitos T , Timo , Animais , Linfócitos B/citologia , Linfócitos B/imunologia , Citometria de Fluxo/história , Citometria de Fluxo/instrumentação , Citometria de Fluxo/métodos , História do Século XX , Camundongos Endogâmicos BALB C , Baço/citologia , Baço/imunologia , Células-Tronco/citologia , Células-Tronco/imunologia , Linfócitos T/citologia , Linfócitos T/imunologia , Timo/citologia , Timo/imunologiaRESUMO
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.