Name: analyzing cellular internalization of nanoparticles and bacteria by multi-spectral imaging flow cytometry
Uploaded: 2012-06-08T13:00:00
Description: Watch this Scientific Journal Video about Analyzing Cellular Internalization of Nanoparticles and Bacteria by Multi-spectral Imaging Flow Cytometry at JoVE.com

作者要感谢金融羚羊的穆里奖（NN00014-06-1-1176）和美国陆军医学研究和装备司令部（批准号W81XWH-09-1-0386和W81XWH-10-1-0806）支持。

1。 RAW 264.7巨细胞培养<ol><li>收获RAW 264.7细胞从他们的保温瓶，当他们到达汇合，他们用细胞刮刀轻轻刮。计数和板块他们到1 5 x 10 5细胞密度/以及在完成0.5毫升Dulbecco的修改鹰中等（cDMEM 24孔细胞培养盘; 10％热灭活胎牛血清（FBS），，2毫米Glutamax 10毫米的肝素钠）在5％CO 2培养箱在37℃孵育过夜。 </li></ol> 2。致病性沙门氏菌沙门伤寒14028转型与文化<ol><li...

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有研究表明，聚（乳酸 - 乙醇酸共聚物（PLGA）或酸酐为基础的可生物降解的纳米粒子可以被用于运载包裹抗原或药物靶细胞，其成效是非常重要的吞噬细胞，这些纳米粒子吸收，从而使定量国际化，设计新颖的纳米给药系统的关键，通过使用这种方法。分析，由多种细胞类型的纳米粒子吸收差可以轻松地分析至目前为止，已量化粒子吸收传统的显微镜和流式细胞仪。然而，各自的替代的方法来研...

<table border="1"><tbody><tr><td> 试剂名称 </td><td> 公司 </td><td> 目录编号 </td><td> 评论 </td></tr><tr><td>的RAW 264.7细胞株</td><td>美国菌种保藏中心（ATCC） </td><td> TIB-71 </td><td></td></tr><tr><td>杜尔贝科的改良Eagle培养基（DMEM培养基） </td><td> cellgro </td><td> 10-013-CV </td><td></td></tr><tr><td>胎牛血清</td><td>亚特兰大生物</td><td>小号11150 </td><td>优质甲级</td></tr><tr><td> glutamax </td><td> Gibco公司</td><td> 35050-061 </td><td></td></tr><tr><td>肝素钠</td><td> Gibco公司</td><td> 15630-080 </td><td></td></tr><tr><td> 24孔板</td><td>卟啉</td><td> 92024 </td><td></td></tr><tr><td>细胞培养瓶</td><td>卟啉</td><td> 90151 </td><td></td></tr><tr><td>细胞刮刀</td><td>卟啉</td><td> 99002 </td><td> 24厘米</td></tr><tr><td>血清型鼠伤寒沙门氏菌 </td><td> ATCC </td><td> 14028 </td><td></td></tr><tr><td> A型肉毒毒素ECM630电的电池机器人</td><td> A型肉毒毒素哈佛仪器</td><td></td><td></td></tr><tr><td>公开资讯观测站</td><td> Fisher Scientific则</td><td> BP308 </td><td></td></tr><tr><td>磷酸盐缓冲液（PBS） </td><td> cellgro </td><td> 21-040-CV </td><td></td></tr><tr><td>超声波液体处理器</td><td>管理信息系统ONIX </td><td>的S-4000 </td><td></td></tr><tr><td>细胞松弛素 - D </td><td> Sigma-Aldrich公司， </td><td> C8273 </td><td></td></tr><tr><td>甲醛</td><td> polysciences </td><td> 04018 </td><td></td></tr><tr><td>洗缓冲区</td><td> 2％热灭活胎牛血清，0.1％叠氮钠的PBS。 </td><td></td><td></td></tr><tr><td>烫发/洗缓冲区</td><td> BD公司</td><td> 554714 </td><td></td></tr><tr><td>清除查看单元帽微管</td><td>西格玛</td><td> T4816 </td><td></td></tr><tr><td>鬼笔660的Alexa Fluor </td><td> Invitrogen公司</td><td> A22285 </td><td></td></tr><tr><td> ImageStream X </td><td> amnis语料库TION </td><td> 100200 </td><td>选项​​：658nm激光，自动进样器</td></tr><tr><td>叠氮化钠</td><td> Fisher Scientific则</td><td>小号227I-500 </td><td></td></tr></tbody></table>

analyzing cellular internalization of nanoparticles and bacteria by multi-spectral imaging flow cytometry

纳米粒子系统已经出现在疫苗交付有价值的工具，通过他们的能力，有效地交付货物，包括蛋白质，抗原提呈细胞1-5。国际化的纳米粒子的抗原提呈细胞（NP），是在封装的抗原产生有效的免疫反应的关键步骤。为了确定在纳米制定影响功能如何变化，我们寻求开发一种高通量，定量实验的协议，该协议是兼容内在的纳米颗粒以及细菌检测。到今天为止，显微镜和流式细胞仪，两个独立的技术，已用于研究纳米粒子的吞噬功能的方法。流式细胞仪的高通量的性质产生了强大的统计数据。然而，由于分辨率低，不能准确地量化与细胞结合纳米粒子内部。显微镜生成高空间分辨率的图像，H贷款本息偿还期中，它是费时，涉及小样本6-8。多光谱成像流式细胞仪（MIFC）是一种新技术，采用显微镜和流式细胞仪同时执行层的核心，通过多种颜色的光谱荧光和亮场成像方面。此功能提供了一个准确的分析荧光信号强度和高速蜂窝不同的结构和功能之间的空间关系。 在此，我们描述了一个方法，利用MIFC特征的细胞群体的内在酸酐纳米粒子或沙门伤寒。我们还描述了制备纳米粒子悬浮液，细胞标记，ImageStream X系统的采集和分析数据的使用理念的应用。我们也证明了一种技术的应用，可以用来区分国际化带够纳米粒子和细菌athways使用的肌动蛋白介导的吞噬作用的抑制剂细胞​​松弛素 - D。

Watch this Scientific Journal Video about Analyzing Cellular Internalization of Nanoparticles and Bacteria by Multi-spectral Imaging Flow Cytometry at JoVE.com

Analyzing Cellular Internalization of Nanoparticles and Bacteria by Multi-spectral Imaging Flow Cytometry

在这篇文章中，我们描述了一种方法，利用多光谱成像流式细胞仪量化酸酐纳米粒子或RAW 264.7细胞细菌的国际化。

由多光谱成像流式细胞仪分析细胞内化纳米粒子和细菌

Nanoparticulate  systems have emerged as valuable tools in vaccine delivery through their  ability to efficiently deliver cargo, including proteins, to ...

Research

JoVE Journal

Bioengineering

Harvesting Murine Alveolar Macrophages and Evaluating Cellular Activation Induced by Polyanhydride Nanoparticles

收获小鼠肺泡巨噬细胞和评价聚酸酐纳米粒子诱导的细胞活化

Biodegradable nanoparticles have emerged as a  versatile platform for the design and implementation of new intranasal vaccines  against respiratory ...

科研

生物工程

Viral Nanoparticles for In vivo Tumor Imaging

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Evaluation of Polymeric Gene Delivery Nanoparticles by Nanoparticle Tracking Analysis and High-throughput Flow Cytometry

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Non-viral gene delivery using polymeric nanoparticles has emerged as an attractive approach for gene therapy to treat genetic diseases1 and as a ...

Wide-field Fluorescent Microscopy and Fluorescent Imaging Flow Cytometry on a Cell-phone

一个手机上的宽视场荧光显微镜和荧光成像流式细胞仪

Fluorescent microscopy and flow cytometry are widely used tools in biomedical research and clinical diagnosis. However these devices are in general ...

Localization and Relative Quantification of Carbon Nanotubes in Cells with Multispectral Imaging Flow Cytometry

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与FRET流式细胞仪灵敏地检测Proteopathic播种活动

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Microwave-driven Synthesis of Iron Oxide Nanoparticles for Fast Detection of Atherosclerosis

动脉粥样硬化的快速检测的氧化铁纳米颗粒微波驱动综合

A fast and reproducible microwave-driven protocol has been developed for the synthesis of neridronate-functionalized nanoparticles. Starting from the ...

Isolation of Primary Murine Retinal Ganglion Cells (RGCs) by Flow Cytometry

Isolation of Primary Murine Retinal Ganglion Cells RGCs by Flow Cytometry

通过流式细胞术分离原代小鼠视网膜神经节细胞（RGC）

Neurodegenerative diseases often have a devastating impact on those affected. Retinal ganglion cell (RGC) loss is implicated in an array of diseases, ...

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化学共轭法在戊型肝炎病毒纳米粒子表面功能化中的应用

Virus-like particles (VLPs) have been used as nanocarriers to display foreign epitopes and/or deliver small molecules in the detection and treatment of ...

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使用光声流细胞测定卵巢癌检测

Many studies suggest that the enumeration of circulating tumor cells (CTCs) may show promise as a prognostic tool for ovarian cancer. Current strategies ...