Name: automated microfluidic blood lysis protocol for enrichment of circulating nucleated cells
Uploaded: 2009-12-31T14:00:00
Description: Watch this Scientific Journal Video about Automated Microfluidic Blood Lysis Protocol for Enrichment of Circulating Nucleated Cells at JoVE.com

研究已经得到了华莱士阁下科尔特基金会早在转化研究事业奖。采血的帮助下添安德鲁斯，小儿血液/肿瘤，。也感谢萨姆豪森博士，流式细胞仪的核心，詹姆斯格雷厄姆布朗癌症中心，运行流式细胞仪检测样品的帮助。

第1部分。微流控设备制造创建一个微流体裂解装置的硅母模使用标准光刻技术与设备，在路易斯维尔洁净室大学[1]。 AutoCAD的（欧特克公司，圣拉菲尔，CA）是用来生成一个透明的面具（Fineline成像，科罗拉多州的斯普林斯，CO），创建渠道的负面副本。从硅掌握使用弹性体聚二甲基硅氧烷（PDMS;道康宁，美联，MI）的软光刻技术的模具制造微流体裂解装置。弹性体与复制的渠道?...

<ol>
	<li>White, W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Clinical+application+of+microfluidic+leukocyte+enrichment+protocol+in+mild+phenotype+sickle+cell+disease+(SCD.">Clinical application of microfluidic leukocyte enrichment protocol in mild phenotype sickle cell disease (SCD.</a> Biomedical Microdevices. 11 (2), 477-483 (2009).</li><li>Sethu, P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Continuous+Flow+Microfluidic+Device+for+Rapid+Erythrocyte+Lysis.">Continuous Flow Microfluidic Device for Rapid Erythrocyte Lysis.</a> Anal. Chem. 76 (21), 6247-6253 (2004).</li></ol>

一个自动化微流体血液溶解协议的报道。值得一提的是在协议的关键步骤。在第P.2.1，重要的是要丢弃的血液小瓶样品中含有成熟的内皮细胞脱落，误报收集。另外，还要确保运行一小时内收集的血液样本。底漆第P.2.2微流体装置，它是重要的，初步摆脱了气泡。此外，应避免引入气泡时附加新的整个协议的针头注射器。 P.2.4节血1 ml注射器灌装时，一定要记住先添加1X PBS和血液，所有同时保持垂?...

automated microfluidic blood lysis protocol for enrichment of circulating nucleated cells

一个自动化微流体血液裂解装置的协议，在本报告中详细介绍。循环有核细胞（CNCS），包括白细胞和内皮细胞，提供一个理想的平台，为个人的免疫状态的更新状态。微流体协议允许无选择性的细胞损失和样品制备变性，由于用户介导的步骤浓缩的CNC。简单地说，协议包括设备制造，样品的采集，设备安装，并通过微腔血。整个器件内的血液迅速去离子水混合，在约10秒50微米× 150微米的微流体通道。在这个时间跨度红细胞裂解由于低渗条件。通道底部的人字形结构，确保充分混合细胞暴露为一个恒定的环境。剩余的单元格返回等渗条件下，在设备的出口，用2％多聚甲醛固定，离心分离CNC的红细胞碎片，并在流缓冲暂停染色和流式细胞仪分析。结果表明清洁流式细胞仪散点图数控保存种群。这种设备和协议的意义数控人口分析疾病的发病机制的研究和理解。因此，自动化，实效性，简单的微流控协议证明。

Watch this Scientific Journal Video about Automated Microfluidic Blood Lysis Protocol for Enrichment of Circulating Nucleated Cells at JoVE.com

Automated Microfluidic Blood Lysis Protocol for Enrichment of Circulating Nucleated Cells

发展循环通过红细胞裂解，以确保高质量的样品无细胞损失隔离外周血有核细胞富集自动化微流体装置。

微流控自动化血液循环核细胞富集的裂解协议

In this report the protocol for an automated microfluidic blood lysis device is detailed. Circulating nucleated cells (CNCs), including leukocytes and ...

Research

JoVE Journal

Biology

Enrichment of NK Cells from Human Blood with the RosetteSep Kit from StemCell Technologies

NK细胞与干细胞技术的RosetteSep套件的人体血液的富集

Natural killer (NK) cells are large granular cytotoxic lymphocytes that belong to the innate immune system and play major roles in fighting against cancer ...

科研

生物学

Microfluidic Co-culture of Epithelial Cells and Bacteria for Investigating Soluble Signal-mediated Interactions

微流体调查可溶性信号介导的相互作用的上皮细胞和细菌的共同文化

The human gastrointestinal (GI) tract is a unique environment in which intestinal epithelial cells and non-pathogenic (commensal) bacteria coexist. It has ...

Isolation and Culture of Pulmonary Endothelial Cells from Neonatal Mice

从新生小鼠的肺血管内皮细胞的分离和培养

Endothelial cells provide a useful research model in many areas of vascular biology. Since its first isolation 1, human umbilical vein endothelial cells ...

Immunofluorescence Analysis of Endogenous and Exogenous Centromere-kinetochore Proteins

内源性的免疫荧光分析及外源着丝粒着丝粒蛋白

"Centromeres" and "kinetochores" refer to the site where chromosomes associate with the spindle during cell division. Direct visualization of ...

Using Microfluidic Devices to Measure Lifespan and Cellular Phenotypes in Single Budding Yeast Cells

使用微流体装置来衡量寿命和细胞表型单芽殖酵母细胞

Budding yeast Saccharomyces cerevisiae is an important model organism in aging research. Genetic studies have revealed many genes with conserved effects ...

A Rapid Automated Protocol for Muscle Fiber Population Analysis in Rat Muscle Cross Sections Using Myosin Heavy Chain Immunohistochemistry

在使用肌球蛋白重链免疫鼠肌肉横断面的快速自动化方案为肌纤维人口分析

Quantification of muscle fiber populations provides a deeper insight into the effects of disease, trauma, and various other influences on skeletal muscle ...

A Graphical User Interface for Software-assisted Tracking of Protein Concentration in Dynamic Cellular Protrusions

软件辅助跟踪动态细胞突变中蛋白质浓度的图形用户界面

Filopodia are dynamic, finger-like cellular protrusions associated with migration and cell-cell communication. In order to better understand the complex ...

A Protocol for Using Gene Set Enrichment Analysis to Identify the Appropriate Animal Model for Translational Research

一个协议来使用基因集富集分析以确定合适的动物模型转化型研究

Recent studies that compared transcriptomic datasets of human diseases with datasets from mouse models using traditional gene-to-gene comparison ...

Isolation of CD4+ T-cells and Analysis of Circulating T-follicular Helper (cTfh) Cell Subsets from Peripheral Blood Using 6-color Flow Cytometry

Isolation of CD4+ T-cells and Analysis of Circulating T-follicular Helper cTfh Cell Subsets from Peripheral Blood Using 6-color Flow Cytometry

cd4+ t 细胞的分离及6色流式细胞仪分析外周血中循环 t-滤泡 (ctfh) 细胞亚群

Aberrant T-follicular helper (Tfh) cell activity is detectable in autoimmune conditions and their presence is associated with clinical outcomes when the ...

Isolation and Enrichment of Human Lung Epithelial Progenitor Cells for Organoid Culture

分离和富集人肺上皮祖细胞用于类器官培养

Epithelial organoid models serve as valuable tools to study the basic biology of an organ system and for disease modeling. When grown as organoids, ...