Name: preparation of single-cell suspensions from mouse spleen with the gentlemacs dissociator
Uploaded: 2008-12-11T00:00:00
Duration: 5 min 18 s
Description: Watch this Scientific Journal Video about Preparation of Single-Cell Suspensions from Mouse Spleen with the gentleMACS Dissociator at JoVE.com

<p class="jove_step">设备简介</p><ol><li>纸巾轻轻分离，使用紫彗星管成可行的单细胞悬液。 </li><li> PBS / BSA EDTA缓冲液是用来分解细胞。缓冲区是准备加入1 BSA的联合解决方案的冲洗液含EDTA autoMACS 20部分。 </li><li>互助委员会预分离过滤器是用来使分离的组织样本中的所有细胞团块，促进成功磁珠分离和后续分析。 </li></ol><p class="jove_step">游离的脾细胞</p><ol><li>全脾disscociated使用gentleMACS Dissociator脾单细胞悬液。 </li><li>要开始的程序，负载​​的缓冲液C管，根据脾脏的质量。一个健康的一个年轻的成年雌性BALB / c小鼠脾重约80-120毫克。这种大规模的每1至2脾脏需要3缓冲液。年龄较大的动物或感染动物的脾脏往往更大，必须在缓冲区权衡。缓冲区卷，必须相应地调整。不要超过10毫升缓冲液。 </li><li>添加解剖脾包含3毫升缓冲到C管。 </li><li>确保了C管帽紧，并重视与第C管dissociator下来。 </li><li>电源了dissociator。 1-2脾脏，选择方案m_spleen_01。该方案将需要56秒钟即可完成。如果脾脏数量较大，使用方案m_spleen_04;这个方案，可用于高达720毫克脾组织。 </li><li>程序完成后本机的脾组织分离成脾细胞悬液。 </li><li>在室温下约30秒300xg离心整个细胞悬液。这将确保转子和定子是在C管的底部收集的所有细胞中解脱出来。 </li><li>通过隔密封在C管帽，使用1000微升pipetter开放取出样品。 </li></ol><p class="jove_step">过滤步骤</p><ol><li>要开始过滤步骤，应用游离组织的预分离的过滤器或用30微米孔径的细胞滤网。超过2脾，过滤网应适合1-2脾脏或50毫升管15毫升管。 </li><li>让细胞悬浮液通过。然后申请5缓冲液冲洗过滤器。 </li><li>现在与细胞悬液，离心管在300xg在室温下，10分钟收集所有的脾细胞，脾细胞沉淀。 </li><li>现在，我们有一个沉淀分离脾，我们必须吸出上清液，重悬细胞沉淀在一个适当的缓冲区数额。 </li></ol><p class="jove_content">使用gentleMACS Dissociator脾虚分解会产生一个可行的脾细胞的比例很高，这是由流动使用MACSQuant分析仪流式细胞仪证明。死细胞染色，碘化丙啶。在所提供的例子，向前侧散射点图显示的准备脾。 </p>

可在所有的协议和数据表<a href="http://www.miltenyibiotec.com"&gt; http://www.miltenyibiotec.com。</a&gt;。

<p>为了避免繁琐而且往往是痛苦的手动的脑机制分离，gentleMACS Dissociator允许游离于组织非常有效地控制和可重复的条件下。组织分解过程是温和的细胞与细胞悬浮液的产量高存活率。 gentleMACS Dissociator与C管结合的目的是准备从各种组织中的单细胞悬液。该仪器配备了预先设定的方案，以分解小鼠脾，肝，肺，或神经组织。方案和协议的范围将不断扩大，以包括额外的组织。 </p>

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<td>gentleMACS&#x2122; Starting Kit</td>
<td>&nbsp;</td>
<td>Miltenyi Biotec</td>
<td>130-093-235</td>
<td><a href="http://www.miltenyibiotec.com/en/PG_709_763_gentleMACS_Dissociator.aspx">http://www.miltenyibiotec.com/en/PG_709_763_gentleMACS_Dissociator.aspx</a></td>
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<td>C Tubes</td>
<td>&nbsp;</td>
<td>Miltenyi Biotec </td>
<td>130-093-237</td>
<td><a href="http://www.miltenyibiotec.com/en/PG_709_765_gentleMACS_C_Tubes.aspx">http://www.miltenyibiotec.com/en/PG_709_765_gentleMACS_C_Tubes.aspx</a></td>
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<td>13 mL Tubes</td>
<td>&nbsp;</td>
<td>Miltenyi Biotec </td>
<td>130-091-596</td>
<td><a href="http://www.miltenyibiotec.com/en/PG_34_751_13_mL_Tubes_for_LS_Columns.aspx">http://www.miltenyibiotec.com/en/PG_34_751_13_mL_Tubes_for_LS_Columns.aspx</a></td>
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<td>autoMACS&#x2122; Rinsing Solution </td>
<td>&nbsp;</td>
<td>Miltenyi Biotec </td>
<td>130-091-222</td>
<td><a href="http://www.miltenyibiotec.com/en/PG_134_155_autoMACS_Rinsing_Solution.aspx">http://www.miltenyibiotec.com/en/PG_134_155_autoMACS_Rinsing_Solution.aspx</a></td>
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<td>MACS&#174; BSA Stock Solution</td>
<td>&nbsp;</td>
<td>Miltenyi Biotec </td>
<td>130-091-376</td>
<td><a href="http://www.miltenyibiotec.com/en/PG_33_295_MACS_BSA_Stock_Solution.aspx">http://www.miltenyibiotec.com/en/PG_33_295_MACS_BSA_Stock_Solution.aspx</a></td>
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<td>Pre-Separation Filters </td>
<td>&nbsp;</td>
<td>Miltenyi Biotec </td>
<td>130-041-407</td>
<td><a href="http://www.miltenyibiotec.com/en/PG_34_247_Pre_Separation_Filters.aspx">http://www.miltenyibiotec.com/en/PG_34_247_Pre_Separation_Filters.aspx</a></td>
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<td>MACSQuant Analyzer</td>
<td>&nbsp;</td>
<td>Miltenyi Biotec </td>
<td>130-092-197</td>
<td>&nbsp;</td>
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<td>Propidium Iodide Solution</td>
<td>&nbsp;</td>
<td>Miltenyi Biotec</td>
<td>130-093-233</td>
<td><a href="http://www.miltenyibiotec.com/en/PG_337_744_Propidium_Iodide_Solution.aspx">http://www.miltenyibiotec.com/en/PG_337_744_Propidium_Iodide_Solution.aspx</a></td>
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preparation of single-cell suspensions from mouse spleen with the gentlemacs dissociator

<p class="jove_content">单细胞悬液，细胞分离，细胞分析，细胞培养实验的一个先决条件。为了避免繁琐而且往往是痛苦的手动的脑机制分离gentleMACS Dissociator允许游离于组织非常有效地控制和可重复的条件下。 gentleMACS Dissociator最佳游离于小鼠脾，结合用户的安全省时和标准化。本视频介绍了如何使用gentleMACS Dissociator，一个自动化的台式设备，可使用特殊管产生可行的细胞悬浮液的机械破坏组织游离于小鼠脾脏。以下分解，脾脏被过滤，离心，并为进一步的应用重悬。</p>

Watch this Scientific Journal Video about Preparation of Single-Cell Suspensions from Mouse Spleen with the gentleMACS Dissociator at JoVE.com

Preparation of Single-Cell Suspensions from Mouse Spleen with the gentleMACS Dissociator

<p class="jove_content">本视频介绍了一个简单的自动组织使用gentleMACS Dissociator准备小鼠脾单细胞悬液的解离，节省时间的技术。</p>

gentleMACS Dissociator从小鼠脾脏单细胞悬浮液的制备

Single-cell suspensions are a prerequisite for experiments in cell separation, cell analysis and cell culture.  To avoid tedious and often painful manual ...

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Research

JoVE Journal

Biology

Standardized Preparation of Single-Cell Suspensions from Mouse Lung Tissue using the gentleMACS Dissociator

标准化的制备小鼠肺组织单细胞悬浮液使用gentleMACS Dissociator

The preparation of single-cell suspensions from tissues is an important prerequisite for many experiments in cellular research. The process of ...

科研

生物学

Generation of Single-Cell Suspensions from Mouse Neural Tissue

从小鼠神经组织产生单细胞悬浮液

Within the nervous system, hundreds of neuronal and glial cell types have been described. Each specific cell type in the brain or spinal cord has a ...

Single-cell Suction Recordings from Mouse Cone Photoreceptors

单从小鼠锥感光细胞吸录音

Rod and cone photoreceptors in the retina are responsible for light detection. In darkness, cyclic nucleotide-gated (CNG) channels in the outer segment ...

Generation of Mice Derived from Induced Pluripotent Stem Cells

诱导多能干细胞来源于小鼠的生成

The production of induced pluripotent stem cells (iPSCs) from somatic cells provides a means to create valuable tools for basic research and may also ...

Simple and Efficient Technique for the Preparation of Testicular Cell Suspensions

简单和高效的睾丸细胞悬液的制备技术

Mammalian testes are very complex organs that contain over 30 different cell types, including somatic testicular cells and different stages of germline ...

Isolation of Endothelial Cells from the Lumen of Mouse Carotid Arteries for Single-Cell Multi-Omics Experiments

Atherosclerosis is an inflammatory disease of the arterial regions exposed to disturbed blood flow (d-flow). D-flow regulates the expression of genes in ...

Real-Time Detection of Reactive Oxygen Species Production in Immune Response in Rice with a Chemiluminescence Assay

用化学发光法实时检测水稻免疫反应中活性氧的产生

Reactive oxygen species (ROS) play vital roles in a variety of biological processes, including the sensing of abiotic and biotic stresses. Upon pathogen ...

Single-Cell Suspension Preparation from Nile Tilapia Intestine for Single-Cell Sequencing

尼罗罗非鱼肠单细胞悬液制备用于单细胞测序

Nile tilapia is one of the most commonly cultured freshwater fish species worldwide and is a widely used research model for aquaculture fish studies. The ...

Isolation and Transcriptome Analysis of Plant Cell Types

植物细胞类型的分离和转录组分析

In multicellular organisms, developmental programming and environmental responses can be highly divergent in different cell types or even within cells, ...

Semi-Automated Isolation of Murine White Adipose Tissue

Semi-Automated Isolation of the Stromal Vascular Fraction from Murine White Adipose Tissue Using a Tissue Dissociator

Semi-Automated Isolation of the Stromal Vascular Fraction from Murine White Adipose Tissue Using a Tissue Dissociator (Video) | JoVE 

使用组织解离器从小鼠白色脂肪组织中半自动分离基质血管部分

The in vitro study of white, brown, and beige adipocyte differentiation enables the investigation of cell-autonomous functions of adipocytes and their ...