Name: monitoring dendritic cell migration using 19f / 1h magnetic resonance imaging
Uploaded: 2013-03-20T13:00:00
Description: Watch this Scientific Journal Video about Monitoring Dendritic Cell Migration using 19F / 1H Magnetic Resonance Imaging at JoVE.com

这项研究是由西南德意志研究联合会（DFG WA 2804）和西南大学教育资助的实验和临床研究中心，最大的一个合作德尔布吕克分子医学中心在柏林的Charité医学院。资助者在研究设计，数据收集和分析，决定发表或准备手稿的作用。我们感谢罗伯特·韦斯特法尔先生在他的技术支持，在我们的实验室实习。

所有动物的程序，必须由当地机构执行前的动物福利委员会批准。于MR测量麻醉和生理监测（体温，呼吸频率）足够水平的必不可少的要求。 1。小鼠骨髓来源的树突状细胞生成<ol><li>提取C57BL / 6小鼠骨髓细胞如前所述，该协议的历史可以追溯到1992年13最初是由拉尔夫·斯坦曼（1943年至二○一一年），树突状细胞的发现者14组 12。 </li><li>简言之...

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这种方法采用19 F / 1ħMRI跟随移动到淋巴结的DC提供了学习的机会， 体内的免疫细胞的迁移模式。树突状细胞是快速迁移的免疫细胞，是能够通过三维结构不紧密粘附的特异性底物17的情况下的机动的很好的例子。虽然所描述的技术的低空间分辨率（μm的范围内）是不与高的分辨率（nm范围内），使用这种技术，可以实现与多光子显微镜，它仍然是可能的，研究细胞?...

细胞在体内的跟踪是在一些生物医药领域的一个重要方面。对于这一点，非侵入性的成像技术，可以选择性地本地化细胞过一段时间是极​​其宝贵的。三维磁共振成像（MRI）的发展之前，不限于显微分析或组织切片的免疫细胞迁移的跟踪。细胞跟踪的帮助下，MRI在过去的几年中取得了巨大的关注，不仅为免疫学研究的免疫细胞在体内的行为，同时也为临床和干细胞研究。在90年代中期，最初的研究氧化铁纳米粒子发起级联发展与MRI追踪细胞。氧化铁颗粒缩短MR弛豫时间（T2 *）标记的细胞，从而导致信号损耗MR图像。氧化铁颗粒已雇用标签的巨噬细胞，少突胶质细胞progenitors 3和许多其它类型的细胞。这些颗粒也被临床被FDA批准用于标记细胞疫苗在黑色素瘤患者4。由于在体内或体外与氧化铁颗粒标记的细胞依赖于缩短的T2 *信号，后者还可以带来体内的易感性相关T2 *的影响，如微出血，铁沉淀物或气泡，因此可能很难识别标记的细胞在体内的来自其他背景T2 *信号灭绝5。 在这篇文章中，我们描述了一种技术用于跟踪在体内的树突状细胞（DC）采用19 F / 1ħ磁共振成像（MRI）。这种细胞追踪技术才被引入几年后，于2005年6 19 F在MRI首次确认申请报告已7。其中一个重要的ADVA氧化铁颗粒细胞标记的19 F超过ntage是低的生物组织中发生的19 F，这使得它可以非常有选择性与基本上无背景图像跟踪细胞。此外，有可能获得从传统的1H核磁共振成像的解剖图像，叠加的19 F标记的细胞从移植的MR信号。因此，19 F / 1ħ核磁共振成像是相当有关的研究在体内细胞迁移。用这种方法研究的细胞都标有19架F-富。综合来自主要由碳原子和氟原子组成的全氟碳（PFC），通常用于制备颗粒。这些化合物是不溶于水和乳化之前， 在体外或体内应用。通常的PFC颗粒大小已受聘于其他群体在体内 19 F-MRI跟踪实验介于100纳米和245纳米6,8-10。然而，我们已经表明，标签与全氟-15 -冠-5 -醚（PFCE）随着粒径（&gt; 560nm的颗粒增加的树突状细胞中的效率）11

<table border="1">
 <tbody>
 <tr>
 <td></td>
 <td></td>
 <td></td>
 <td>REAGENTS</td>
 </tr>
 <tr>
 <td>C57BL/6 mice</td>
 <td>Charles River, Berlin</td>
 <td></td>
 <td></td>
 </tr>
 <tr>
 <td>RPMI</td>
 <td>Gibco</td>
 <td>21875-091</td>
 <td></td>
 </tr>
 <tr>
 <td>FBS Superior</td>
 <td>Biochrom AG</td>
 <td>S 0615</td>
 <td></td>
 </tr>
 <tr>
 <td>HEPES</td>
 <td>Gibco-Invitrogen</td>
 <td>15630-056</td>
 <td></td>
 </tr>
 <tr>
 <td>Penicillin-Streptomycin</td>
 <td>Gibco</td>
 <td>15140-122</td>
 <td></td>
 </tr>
 <tr>
 <td>L-glutamine</td>
 <td>Gibco</td>
 <td>25030-024</td>
 <td></td>
 </tr>
 <tr>
 <td>Dulbecco's PBS</td>
 <td>Sigma Aldrich</td>
 <td>D8662</td>
 <td></td>
 </tr>
 <tr>
 <td>PFA</td>
 <td>Santa Cruz</td>
 <td>sc-281692</td>
 <td></td>
 </tr>
 <tr>
 <td>Perfluoro-15-crown-5-ether </td>
 <td>ChemPur</td>
 <td>391-1996</td>
 <td></td>
 </tr>
 <tr>
 <td>Pluronic F-68 </td>
 <td>Sigma Aldrich</td>
 <td>P5556</td>
 <td></td>
 </tr>
 <tr>
 <td>Petri dishes (35 x 10 mm)</td>
 <td>VWR, Germany</td>
 <td>391-1996</td>
 <td></td>
 </tr>
 <tr>
 <td>27 &frac12; G syringes</td>
 <td>VWR, Germany</td>
 <td>612-0151</td>
 <td></td>
 </tr>
 <tr>
 <td>Nylon cell strainers (100 &mu;m mesh) </td>
 <td>VWR, Germany</td>
 <td>734-0004</td>
 <td></td>
 </tr>
 <tr>
 <td>NMR tubes </td>
 <td>VWR, Germany</td>
 <td>634-0461</td>
 <td></td>
 </tr>
 <tr>
 <td></td>
 <td></td>
 <td></td>
 <td>EQUIPMENT</td>
 </tr>
 <tr>
 <td>Dissection tools</td>
 <td>FST</td>
 <td></td>
 <td></td>
 </tr>
 <tr>
 <td>CO2 incubator </td>
 <td>Binder</td>
 <td></td>
 <td></td>
 </tr>
 <tr>
 <td>Small animal MR system</td>
 <td>Bruker Biospin</td>
 <td></td>
 <td>9.4T BioSpec 94/20 USR, ParaVision Acquisition and Processing Software</td>
 </tr>
 <tr>
 <td>1H/19F dual-tunable volume RF coil </td>
 <td>Rapid Biomed, W&uuml;rzburg, Germany</td>
 <td></td>
 <td>35 mm inner diameter, 50 mm length</td>
 </tr>
 <tr>
 <td>19F spectroscopy coil </td>
 <td>in-house</td>
 <td></td>
 <td>tune/match loop coil, 4 turns, inner diameter 5 mm, 10 mm long, two capacitors for tuning and matching</td>
 </tr>
 <tr>
 <td>Isoflurane inhalation system</td>
 <td>F&ouml;hr Medical Instruments GmbH</td>
 <td></td>
 <td></td>
 </tr>
 <tr>
 <td>Animal monitoring system Model 1025</td>
 <td>SA Instruments Inc., New York, USA</td>
 <td></td>
 <td></td>
 </tr>
 </tbody>
</table>

monitoring dendritic cell migration using 19f / 1h magnetic resonance imaging

如磁共振成像（MRI）的非侵入性的成像方式的不断进步，极大地提高了我们的学习能力，在生物体的生理或病理过程。 MRI也证明是一个有价值的工具捕捉移植细胞在体内 。初始小区标记策略用于MRI使用的造影剂的影响的MR弛豫时间（T1，T2，T2 *）和引起增强（T1）或耗尽（T2 *）标记的细胞存在的信号。 T2 *增强剂，如超小型氧化铁剂（USPIO）已经被用来研究细胞迁移和一些人也已被FDA批准用于临床应用。 T2 *剂的缺点是难以区分从其他文物，如血块，微出血或气泡标记细胞的信号灭绝创建的。在这篇文章中，我们描述了一个新兴的技术跟踪细胞在体内基于氟（第19楼）丰富的颗粒细胞标记。这些颗粒的制备是通过乳化的全氟化碳（PFC）的化合物，然后用的标签细胞，其后，可以通过19 F核磁共振成像成像。的PFCs的重要的优点包括（i） 在体内的碳结合的19 F 在体内 ，然后通过19 F磁共振波谱产生背景图像和量化的细胞信号的完整的细胞选择性强（ⅱ）的可能性的情况下进行细胞跟踪。

18〜24小时后，皮内的应用程序，19 F-标记的树突状细胞（DC）迁移到排水的腘淋巴结。可以理解的DC，通过到排水politeal的淋巴结的淋巴管的运动，通过叠加的1 H 19 F DC图像（ 图2A）的解剖图像。我们以前曾报道这些细胞在体内的迁移，以及19 F-粒度对DC免疫生物学，包括吸收效率11的影响。为了量化的DC迁移到淋巴结的程度，我们提取引流淋巴结?...

Watch this Scientific Journal Video about Monitoring Dendritic Cell Migration using 19F / 1H Magnetic Resonance Imaging at JoVE.com

Monitoring Dendritic Cell Migration using 19F / 1H Magnetic Resonance Imaging

细胞利用磁共振成像跟踪，在过去几年里取得了显着的关注。本协议描述标签的树突状细胞与氟（<sup&gt; 19</sup&gt; F）的丰富的颗粒，这些细胞在体内的应用程序，和监测他们迁移到引流淋巴结的程度<sup&gt; 19</sup&gt; F /<sup&gt; 1</sup&gt; H MRI和<sup&gt; 19</sup&gt; F刘健。

监测树突状细胞迁移<sup&gt; 19</sup&gt; F /<sup&gt; 1</sup&gt; H磁共振成像

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白细胞的归位和迁移的非侵入性的成像在体内</em

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样品制备<em&gt;结核分枝杆菌</em提取物对核磁共振的代谢组学研究

Mycobacterium tuberculosis is a major cause of mortality in human beings on a global scale. The emergence of both multi- (MDR) and extensively-(XDR) ...

Analysis of Pulmonary Dendritic Cell Maturation and Migration during Allergic Airway Inflammation

过敏性气道炎症在肺树突状细胞的成熟和迁移分析

Dendritic cells (DCs) are the key players involved in initiation of adaptive immune response by activating antigen-specific T cells. DCs are present in ...

Generation of a Novel Dendritic-cell Vaccine Using Melanoma and Squamous Cancer Stem Cells

使用黑色素瘤和鳞状癌干细胞生成新型支气干细胞疫苗

We identified cancer stem cell (CSC)-enriched populations from murine melanoma D5 syngeneic to C57BL/6 mice and the squamous cancer SCC7 syngeneic to C3H ...

Imaging CD4 T Cell Interstitial Migration in the Inflamed Dermis

成像的CD4 T细胞间质迁移发炎真皮

The ability of CD4 T cells to carry out effector functions is dependent upon the rapid and efficient migration of these cells in inflamed peripheral ...

A CFSE-based Assay to Study the Migration of Murine Skin Dendritic Cells into Draining Lymph Nodes During Infection with Mycobacterium bovis Bacille Calmette-Gu&#233;rin

A CFSE-based Assay to Study the Migration of Murine Skin Dendritic Cells into Draining Lymph Nodes During Infection with Mycobacterium bovis Bacille Calmette-GuÃƒÆ’Ã†'Ãƒâ€šÃ‚Â©rin

基于CFSE-试验鼠皮肤树突状细胞的迁移习进引流淋巴结在与感染<em&gt;牛分枝杆菌</em&gt;卡介苗

Dendritic cells (DCs) are important for initiating immune responses, in part through their ability to acquire and shuttle antigen to the draining lymph ...

Measurement of T Cell Alloreactivity Using Imaging Flow Cytometry

使用成像流式细胞仪T细胞同种异体反应性测定

The measurement of immunological reactivity to donor antigens in transplant recipients is likely to be crucial for the successful reduction or withdrawal ...