Name: in situ quantification of pancreatic beta-cell mass in mice
Uploaded: 2010-06-07T17:00:00
Description: Watch this Scientific Journal Video about In situ Quantification of Pancreatic Beta-cell Mass in Mice at JoVE.com

支持这项研究是由美国公众卫生服务补助DK - 081527，DK - 072473和DK - 20595芝加哥糖尿病研究和培训中心（动物模型核心）的大学，Kovler家庭基金会的礼物。

第1部分：试样制备<ol><li>下解剖镜下放置鼠标，整个胰腺仍然附着与十二指肠和脾脏中删​​除，并放在一个预先称重的载玻片机关。取出沿侧切割结缔组织，它必然胰腺十二指肠。剪切和删除对胰腺的脾和多余的脂肪，直到胰腺是幻灯片上的完全隔离。 （胰腺脂肪是从胰腺组织的颜色稍深和Tanner的区别，其白色区分开来。） </li><li>将盖玻片（大盖玻璃50x75mm;标准盖玻片25x75mm）在胰腺，使其完...

<ol>
	<li>Miller, K., Kim, A., Kilimnik, G., Moka, U., Jo, J., Periwal, V., Hara, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Islet+formation+during+the+neonatal+development+in+mice.">Islet formation during the neonatal development in mice.</a> PloS One. 4, E7739-E7739 (2009).</li><li>Kilimnik, G., Kim, A., Jo, J., Miller, K., Hara, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Quantification+of+pancreatic+islet+distribution+in+situ+in+mice.">Quantification of pancreatic islet distribution in situ in mice.</a> Am J Physiol Endocrinol Metab. 297, E1331-E1338 (2009).</li><li>Hara, M., Wang, X., Kawamura, T., Bindokas, V. P., Dizon, R. F., Alcoser, S. Y., Magnuson, M. A., Bell, G. I. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Transgenic+mice+with+green+fluorescent+protein-labeled+pancreatic+beta+-cells.">Transgenic mice with green fluorescent protein-labeled pancreatic beta -cells.</a> Am J Physiol Endocrinol Metab. 284, E177-E183 (2003).</li><li>Hara, M., Dizon, R. F., Glick, B. S., Lee, C. S., Kaestner, K. H., Piston, D. W., Bindokas, V. P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Imaging+pancreatic+beta-cells+in+the+intact+pancreas.">Imaging pancreatic beta-cells in the intact pancreas.</a> Am J Physiol Endocrinol Metab. 290, E1041-E1047 (2006).</li></ol>

完整的胰腺中的胰岛细胞和β细胞集群的虚拟切片图像分析提供了一个大规模的看法，整个小岛分布。发展，随着时间的推移，总胰岛分布的变化，因此可以研究和比较。在我们的胰岛形成（1）分析表明了这样一个例子。在不同时间点（P1 - P21）的综合分析表明新生小鼠的胰腺，胰岛裂变形成。虽然β-细胞的增殖与小鼠的对数正态分布的概率密度函数从P1 - P10适合从P12开始，胰岛分布向左偏离距离...

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		<th>Catalogue Number</th>
		<th>Comment</th>
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<td>Forceps</td>
<td>&nbsp;</td>
<td>Miltex</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
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<td>Scissors</td>
<td>&nbsp;</td>
<td>F.S.T</td>
<td>14001-13</td>
<td>01n2</td>
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<td>Scissors</td>
<td>&nbsp;</td>
<td>F.S.T</td>
<td>14072-10</td>
<td>02s5	</td>
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<tr>
<td>Large glass slide</td>
<td>&nbsp;</td>
<td>Electron Microscopy Sciences</td>
<td>71862-01</td>
<td>75x51x1.2mm</td>
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<tr>
<td>Large cover glass</td>
<td>&nbsp;</td>
<td>Ted Pella, Inc</td>
<td>260462</td>
<td>50x75mm</td>
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<tr>
<td>Glass Slide</td>
<td>&nbsp;</td>
<td>Fisher Scientific</td>
<td>12-550-15</td>
<td>25x75x1.0mm</td>
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<tr>
<td>Cover glass</td>
<td>&nbsp;</td>
<td>Fisher Scientific</td>
<td>12-458-5M</td>
<td>75x25mm</td>
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<tr>
<td>Cover glass</td>
<td>&nbsp;</td>
<td>Erie Scientific</td>
<td>&nbsp;</td>
<td>25mm circle</td>
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<td>Fluorescent microscope</td>
<td>&nbsp;</td>
<td>Olympus IX-81</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
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<td>Dissection microscope</td>
<td>&nbsp;</td>
<td>Olympus SZX12</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
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<td>Stereo Investigator</td>
<td>&nbsp;</td>
<td>MicroBrightField</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
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<td>MIP-GFP mice</td>
<td>&nbsp;</td>
<td>Jackson Laboratory</td>
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in situ quantification of pancreatic beta-cell mass in mice

跟踪在健康和疾病的特定细胞群的变化是生物医学研究的一个重要目标。监测胰腺β细胞增殖和胰岛生长的过程中是特别具有挑战性。我们已经开发出一种方法来捕获完整的胰腺β细胞的分布与荧光标记的β细胞ImageJ（/ rsb.info.nih.gov / IJ）编写的宏转基因小鼠。胰腺解剖和组织清算，作为一个虚拟切片，捕获整个胰腺后，GFP标记的β细胞的研究。分析包括定量β细胞总面积，胰岛数量和大小分布参考具体参数和每个胰岛β细胞小群的位置。可以绘制整个小岛分布在三个层面，从每个胰岛的大小和形状的分布信息，允许一个整体β细胞面积的变化一目了然的定量和定性的比较。

Watch this Scientific Journal Video about In situ Quantification of Pancreatic Beta-cell Mass in Mice at JoVE.com

In situ Quantification of Pancreatic Beta-cell Mass in Mice

以下协议概述了虚拟切片成像过程中，胰腺癌的夹层和随后所有GFP标记在整个胰腺β细胞的定量。

在小鼠胰腺β-细胞团的原位定量

Tracing changes of specific cell populations in health and disease is an important goal of biomedical research. The process of monitoring pancreatic ...

Research

JoVE Journal

Biology

In situ Quantification of Pancreatic Beta-cell Mass in Mice

Transplantation of Pancreatic Islets Into the Kidney Capsule of Diabetic Mice

补肾胶囊对糖尿病小鼠的胰岛移植

Our protocol was developed to cleanly and easily deliver islets or cells under the kidney capsule of diabetic or normal mice.  We found that it was easier ...

科研

生物学

Quantification of &gamma;H2AX Foci in Response to Ionising Radiation

Quantification of ÃƒÅ½Ã‚Â³H2AX Foci in Response to Ionising Radiation

电离辐射的反应定量的γH2AX灶

DNA double-strand breaks (DSBs), which are induced by either endogenous metabolic processes or by exogenous sources, are one of the most critical DNA ...

Computer-assisted Large-scale Visualization and Quantification of Pancreatic Islet Mass, Size Distribution and Architecture

计算机辅助大型胰岛质量，粒度分布和结构的可视化和量化

The pancreatic islet is a unique micro-organ composed of several hormone secreting endocrine cells such as beta-cells (insulin), alpha-cells (glucagon), ...

Quantification of Proteins Using Peptide Immunoaffinity Enrichment Coupled with Mass Spectrometry

加上质谱肽免疫亲和富集的蛋白质定量

There is a great need for quantitative assays in measuring proteins. Traditional sandwich immunoassays, largely considered the gold standard in ...

Evaluation of Muscle Function of the Extensor Digitorum Longus Muscle Ex vivo and Tibialis Anterior Muscle In situ in Mice

Evaluation of Muscle Function of the Extensor Digitorum Longus Muscle Ex vivo and Tibialis Anterior Muscle In situ in Mice

伸趾长肌的肌肉功能评价<em&gt;体外</em和胫骨前肌<em在原位</em在小鼠

Body  movements are mainly provided by mechanical function of skeletal muscle. Skeletal  muscle is composed of numerous bundles of myofibers that are ...

Visualization and Analysis of mRNA Molecules Using Fluorescence In Situ Hybridization in Saccharomyces cerevisiae

Visualization and Analysis of mRNA Molecules Using Fluorescence In Situ Hybridization in Saccharomyces cerevisiae

可视化和分析的mRNA分子，使用荧光<em&gt;在原位</em&gt;杂交<em&gt;酿酒酵母（Saccharomyces cerevisiae）</em

The Fluorescence in situ Hybridization (FISH) method allows one to detect nucleic acids in the native cellular environment. Here we provide a protocol for ...

Selected Reaction Monitoring Mass Spectrometry for Absolute Protein Quantification

选择反应监测质谱法进行绝对定量蛋白质

Absolute quantification of target proteins within complex biological samples is critical to a wide range of research and clinical applications. This ...

High Resolution Quantification of Crystalline Cellulose Accumulation in Arabidopsis Roots to Monitor Tissue-specific Cell Wall Modifications

High Resolution Quantification of Crystalline Cellulose Accumulation in Arabidopsis Roots to Monitor Tissue-specific Cell Wall Modifications

结晶纤维素积累高分辨率的量化<em&gt;拟南芥</em&gt;根监视特定组织细胞壁修改

Plant cells are surrounded by a cell wall, the composition of which determines their final size and shape. The cell wall is composed of a complex matrix ...

Observation and Quantification of Telomere and Repetitive Sequences Using Fluorescence In Situ Hybridization (FISH) with PNA Probes in Caenorhabditis elegans

Observation and Quantification of Telomere and Repetitive Sequences Using Fluorescence In Situ Hybridization FISH with PNA Probes in Caenorhabditis elegans

观察与端粒的定量和重复序列用荧光<em&gt;原位</em&gt;杂交（FISH）与PNA探针<em&gt;秀丽隐杆线虫</em

Telomere is a ribonucleoprotein structure that protects chromosomal ends from aberrant fusion and degradation. Telomere length is maintained by telomerase ...

Analysis of Beta-cell Function Using Single-cell Resolution Calcium Imaging in Zebrafish Islets

斑马鱼胰岛单细胞分辨率钙成像对β细胞功能的分析

Pancreatic beta-cells respond to increasing blood glucose concentrations by secreting the hormone insulin. The dysfunction of beta-cells leads to ...