Name: labeling stem cells with fluorescent dyes for non-invasive detection with optical imaging
Uploaded: 2008-04-02T18:00:00
Description: Watch this Scientific Journal Video about Labeling Stem Cells with Fluorescent Dyes for non-invasive Detection with Optical Imaging at JoVE.com

<p>这个项目是一个莱昂J.塔尔从加州再生医学研究所的种子批。托比亚斯亨宁是由德国研究协会（DFG，何4578/1-2）津贴一个研究。我们要感谢他对人类胚胎干细胞培养的意见的华尼托Meneses。 </p>

<p class="jove_step">间质干细胞的标记与荧光染料</p><ol><li>要开始标签间质干细胞的过程，trypsinize和细胞计数，以获得与一个细胞的定义暂停。 </li><li>的孵化器中取出细胞，吸出含有被标记的细胞培养瓶旧媒体</li><li> 10毫升的Mg / Ca无PBS清洗细胞。吸出的PBS。镁和钙会抑制胰蛋白酶，因此我们用PBS没有这些。 </li><li>加入预热的0.05％胰蛋白酶，为T75烧瓶中，我们使用5毫升。确保覆盖整个表面的容量瓶。 </li><li>在37 ° C?...

<ol>
	<li>Daldrup-Link, H. E., Rudelius, M., Metz, S., Piontek, G., Settles, M., Pichler, B., Heinzmann, U., Weinmann, H. J., Schlegel, J., Link, T. M., Rummeny, E. J., Oostendorp, R. A. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Stem+cell+tracking+with+Gadophrin-2+--+a+bifunctional+contrast+agent+for+MR+imaging,+optical+imaging+and+fluorescence+microscopy.">Stem cell tracking with Gadophrin-2 -- a bifunctional contrast agent for MR imaging, optical imaging and fluorescence microscopy.</a> <em style='font-style: italic'>Eur J Nucl Med Mol Imaging</em>. <b>31</b>, 1312-1321 (2004).</li><li>Simon, G. H., Daldrup-Link, H. E., Kau, J., Metz, S., Schlegel, J., Piontek, G., Saborowski, O., Demos, S., Duyster, J., Pichler, B. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Optical+imaging+of+experimental+arthritis+using+allogeneic+leukocytes+labeled+with+a+near-infrared+fluorescent+probe.">Optical imaging of experimental arthritis using allogeneic leukocytes labeled with a near-infrared fluorescent probe.</a> <em style='font-style: italic'>Eur J Nucl Med &amp; Mol Imaging</em>. <b>33</b>, 998-1006 (2006).</li><li>Sutton, E., Henning, T., Pichler, B., Bremer, C., Daldrup-Link, H. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Cell+Tracking+with+Optical+imaging.">Cell Tracking with Optical imaging.</a> <em style='font-style: italic'>Eur Radiol</em>. <b>2</b>, 350-357 (2003).</li><li>Funovics, M. A., Alencar, H., Su, H. S., Khazaie, K., Weissleder, R., Mahmood, U. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Miniaturized+multichannel+near+infrared+endoscope+for+mouse+imaging.">Miniaturized multichannel near infrared endoscope for mouse imaging.</a> <em style='font-style: italic'>Mol Imaging</em>. <b>2</b>, 350-357 (2003).</li></ol>

<p> Oi是一个相对较新的成像技术，荧光检测的基础上。 Oi是示踪剂成像技术的敏感，但不与任何辐射暴露关联。其他投资提供跟踪细胞的非侵入性的和重复的一个有效手段，从而提供了了解细胞迁移到目标站点。该技术的一个主要限制是有限的荧光探针在体内的组织渗透。因此，在深部组织的示踪积累，多约5-10厘米的皮肤表面，可能不会被发现。目前临床上的应用是有限的肤浅的技术，如内镜，心血管疾病和视网...

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		<th>Material Name</th>
		<th>Type</th>
		<th>Company</th>
		<th>Catalogue Number</th>
		<th>Comment</th>
		</tr>
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<tr>
<td>Indocyanine Green (IR-125)</td>
<td>Reagent</td>
<td>Fisher Scientific</td>
<td>AC41254-1000 </td>
<td>&nbsp;</td>
<tr>
<td>DMSO</td>
<td>Reagent</td>
<td>Sigma</td>
<td>D-2650 </td>
<td>&nbsp;</td>
<tr>
<td>D-MEM High Glucose </td>
<td>Reagent</td>
<td>Sigma</td>
<td>D5648</td>
<td>&nbsp;</td>
<tr>
<td>Gelatin</td>
<td>Reagent</td>
<td>Sigma</td>
<td>G1890</td>
<td>Dilute to final concentration of 0.1% in PBS</td>
</tr>
<tr>
<td>PBS, Mg and Ca free</td>
<td>Reagent</td>
<td>GIBCO </td>
<td>14190-144</td>
<td>&nbsp;</td>
<tr>
<td>Trypsin-EDTA 0.05% </td>
<td>Reagent</td>
<td>Invitrogen</td>
<td>25300-120</td>
<td>&nbsp;</td>
<tr>
<td>Trypsin-EDTA 0.25% </td>
<td>Reagent</td>
<td>Invitrogen</td>
<td>25200-114	</td>
<td>&nbsp;</td>
<tr>
<td>FCS, characterized</td>
<td>Reagent</td>
<td>Hyclone</td>
<td>SH30071.03</td>
<td>&nbsp;</td>
<tr>
<td>Cell Strainer (40 &#x03BC;m Nylon)</td>
<td>Reagent</td>
<td>BD Falcon</td>
<td>352340</td>
<td>&nbsp;</td>
<tr>
<td>DiD</td>
<td>Reagent</td>
<td>Molecular Probes</td>
<td>V-22887</td>
<td>
</td>
</tr>
<tr>
<td>Knockout DMEM</td>
<td>Reagent</td>
<td>GIBCO</td>
<td>10829018</td>
<td>&nbsp;</td>
<tr>
<td>Knockout Serum Replacer</td>
<td>Reagent</td>
<td>GIBCO</td>
<td>10828028</td>
<td>
</td>
</tr>
<tr>
<td>b-Mercaptoethanol </td>
<td>Reagent</td>
<td>Sigma</td>
<td>7522</td>
<td>&nbsp;</td>
<tr>
<td>Non-essential Amino Acids </td>
<td>Reagent</td>
<td>GIBCO</td>
<td>11140050</td>
<td>&nbsp;</td>
<tr>
<td>FGF-2</td>
<td>Reagent</td>
<td>R&D Systems </td>
<td> 233-FB-025</td>
<td>&nbsp;</td>
<tr>
<td>Glutamine 200mM </td>
<td>Reagent</td>
<td>GIBCO</td>
<td>25030081</td>
<td>&nbsp;</td>
<tr>
<td>Penicillin/Streptomycin</td>
<td>Reagent</td>
<td>GIBCO</td>
<td>15140-122</td>
<td>&nbsp;</td>
<tr>
<td>Protamine Sulfate</td>
<td>Reagent</td>
<td>American Pharmaceutical Partners Inc.</td>
<td>&nbsp;</td>
<td>&nbsp;</td>		</tbody>
		</table>
		</div>
	

labeling stem cells with fluorescent dyes for non-invasive detection with optical imaging

<p class="jove_content">光学成像（OI），是一个方便，快捷，廉价的新的干细胞治疗的体内监测的工具。该技术是基于体内的干细胞用荧光染料，随后静脉注射标记的细胞，在特定的靶器官或病理积累的可视化标签。所提出的技术演示我们如何由简单的孵化与亲脂性荧光染料标签人类间质干细胞（HMSC）（C67H103CIN2O3S），我们如何标签人类胚胎干细胞（胚胎干细胞），美国FDA批准的荧光染料的吲哚菁绿（ICG）。吸收机制是通过坚持和推广整个磷脂细胞膜双层lypophilic染料。通常标记效率提高，如果细胞培养，而不是在含血清的介质中孵化的无血清培养基中的染料。此外，除了转染剂硫酸鱼精蛋白显着提高造影剂的吸收。</p>

Watch this Scientific Journal Video about Labeling Stem Cells with Fluorescent Dyes for non-invasive Detection with Optical Imaging at JoVE.com

Labeling Stem Cells with Fluorescent Dyes for non-invasive Detection with Optical Imaging

与非侵入性检测荧光染料标记的干细胞与光学成像

<p class="jove_content">该视频显示技术，人类胚胎干细胞和骨髓基质干细胞与荧光染料标签。这种技术可以用于移植的干细胞在体内追踪与光学成像和荧光显微镜观察组织病理学的相关性。</p>

Optical imaging (OI) is an easy, fast and inexpensive tool for in vivo monitoring of new stem cell based therapies. The technique is based on ex vivo ...

Research

JoVE Journal

Biology

Labeling hESCs and hMSCs with Iron Oxide Nanoparticles for Non-Invasive in vivo Tracking with MR Imaging

氧化铁标记的胚胎干细胞和的hMSCs纳米粒子在体内的跟踪与非侵入性磁共振成像

In recent years, stem cell research has led to a better understanding of developmental biology, various diseases and its potential impact on regenerative ...

科研

生物学

Non-invasive 3D-Visualization with Sub-micron Resolution Using Synchrotron-X-ray-tomography

非侵入性的三维可视化与亚微米分辨率使用同步加速器X射线断层扫描

Little is known about the internal organization of many micro-arthropods with body sizes below 1 mm. The reasons for that are the small size and the hard ...

In vitro Labeling of Human Embryonic Stem Cells for Magnetic Resonance Imaging

在人类胚胎干细胞体外培养的标签用于磁共振成像

Human embryonic stem cells (hESC) have demonstrated the ability to restore the injured myocardium. Magnetic resonance imaging (MRI) has emerged as one of ...

Proper Care and Cleaning of the Microscope

适当的照顾和清洁的显微镜

Keeping the microscope optics clean is important for high-quality imaging. Dust, fingerprints, excess immersion oil, or mounting medium on or in a ...

Phase Contrast and Differential Interference Contrast (DIC) Microscopy

Phase Contrast and Differential Interference Contrast DIC Microscopy

相衬，微分干涉对比（DIC）显微镜

Phase-contrast microscopy is often used to produce contrast for transparent, non light-absorbing, biological specimens. The technique was discovered by ...

Visualizing the Live <em>Drosophila</em> Glial-neuromuscular Junction with Fluorescent Dyes

Visualizing the Live Drosophila Glial-neuromuscular Junction with Fluorescent Dyes

可视化的Live<em>果蝇</em>胶质细胞神经​​肌肉交界处，与荧光染料

Our project identified GFP labeled glial structures at the developing larval fly neuromuscular synapse. To look at development of live glial-nerve-muscle ...

Fluorescent Labeling of <em>Drosophila</em> Heart Structures

Fluorescent Labeling of Drosophila Heart Structures

荧光标记的<em>果蝇</em>心结构

The Drosophila melanogaster dorsal vessel, or heart, is a tubular structure comprised of a single layer of contractile cardiomyocytes, pericardial cells ...

Lineage Labeling of Zebrafish Cells with Laser Uncagable Fluorescein Dextran

天堂斑马鱼的细胞标记与激光Uncagable荧光葡聚糖

A central problem in developmental biology is to deduce the origin of the myriad cell types present in vertebrates as they arise from undifferentiated ...

Fluorescence Imaging with One-nanometer Accuracy (FIONA)

Fluorescence Imaging with One-nanometer Accuracy FIONA

荧光成像与一纳米精度（FIONA）

Fluorescence imaging with one-nanometer accuracy (FIONA) is a simple but useful technique for localizing single fluorophores with nanometer precision in ...

Detection of Modified Forms of Cytosine Using Sensitive Immunohistochemistry

胞嘧啶修饰形式使用敏感免疫组化检测

Methylation of cytosine bases (5-methylcytosine, 5mC) occurring in vertebrate genomes is usually associated with transcriptional silencing. ...