Name: quantifying yeast chronological life span by outgrowth of aged cells
Uploaded: 2009-05-06T00:00:00
Description: Watch this Scientific Journal Video about Quantifying Yeast Chronological Life Span by Outgrowth of Aged Cells at JoVE.com

这项工作是支持由美国国立卫生研究院资助1R21AG031965 - 01A1。 Mk为中老年埃利森医学基金会新学者。

第1部分：老龄化的文化准备<ol><li>到YEPD培养基琼脂平板冻结的股票（1％酵母提取物，细菌用2％的蛋白胨，2％琼脂，2％葡萄糖）利息数株。 </li><li>在30˚C的48小时或直到出现单菌落培养的细胞。 </li><li>选入YEPD液体培养基在试管中的5毫升（1％酵母提取物，细菌用的蛋白胨2％，2％葡萄糖）单菌落，接种。 </li><li>拓展文化30一夜之间˚C，而保持恒定的搅拌，使用一个振动筛或滚鼓。 </li><li> 50μL过...

<ol>
	<li>Steinkraus, K. A., Kaeberlein, M., Kennedy, B. K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Replicative+aging+in+yeast:+the+means+to+the+end.">Replicative aging in yeast: the means to the end.</a> Annu Rev Cell Dev Biol. 24, 29 (2008).</li><li>Kaeberlein, M., Conn, P. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Handbook of models for human aging. , 109 (2006).</li><li>Fabrizio, P., Longo, V. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+chronological+life+span+of+Saccharomyces+cerevisiae.">The chronological life span of Saccharomyces cerevisiae.</a> Aging Cell. 2 (2), 73 (2003).</li><li>Murakami, C. J., Burtner, C. R., Kennedy, B. K., Kaeberlein, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+method+for+high-throughput+quantitative+analysis+of+yeast+chronological+life+span.">A method for high-throughput quantitative analysis of yeast chronological life span.</a> J Gerontol A Biol Sci Med Sci. 63 (2), 113 (2008).</li><li>Piper, P. W., Harris, N. L., MacLean, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Preadaptation+to+efficient+respiratory+maintenance+is+essential+both+for+maximal+longevity+and+the+retention+of+replicative+potential+in+chronologically+ageing+yeast.">Preadaptation to efficient respiratory maintenance is essential both for maximal longevity and the retention of replicative potential in chronologically ageing yeast.</a> Mech Ageing Dev. 127 (9), 733 (2006).</li><li>Fabrizio, P., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Superoxide+is+a+mediator+of+an+altruistic+aging+program+in+Saccharomyces+cerevisiae.">Superoxide is a mediator of an altruistic aging program in Saccharomyces cerevisiae.</a> J Cell Biol. 166 (7), 1055 (2004).</li></ol>

高通量的时间顺序寿命试验，这里描述的是一个量化的菌株具有高准确度和精密度的大量老化潜力的有效方法。这种方法比传统方法主要事先确定计数菌落形成单位（如见3）的生存是一个摇床/孵化器/板读取装置，如使用的BIOSCREEN彗星MBR机获取高分辨率的生长曲线每个年龄点。在直接比较低吞吐量，按时间顺序寿命试验，这种方法已被证明，以达到可比性（或更好）的精度，同时可以...

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		<th>Catalogue Number</th>
		<th>Comment</th>
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<td>Bacto Peptone</td>
<td>Reagent</td>
<td>BD</td>
<td>211677</td>
<td>&nbsp;</td>
<tr>
<td>Bacto Yeast Extract</td>
<td>Reagent</td>
<td>BD</td>
<td>288620</td>
<td>&nbsp;</td>
<tr>
<td>Difco Agar</td>
<td>Reagent</td>
<td>BD</td>
<td>214530</td>
<td>&nbsp;</td>
<tr>
<td>Yeast Nitrogen Base w/o A.A. and A.S.</td>
<td>Reagent</td>
<td>MidSci</td>
<td>J630-500G</td>
<td>&nbsp;</td>
<tr>
<td>Amino Acids</td>
<td>Reagent</td>
<td>Sigma</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<tr>
<td>Ammonium Sulfate</td>
<td>Reagent</td>
<td>Spectrum</td>
<td>AM185</td>
<td>&nbsp;</td>
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<td>Dextrose</td>
<td>Reagent</td>
<td>Fisher</td>
<td>D16-10</td>
<td>&nbsp;</td>
<tr>
<td>Bioscreen C MBR machine</td>
<td>Tool</td>
<td>Growth Curves USA</td>
<td>5101370</td>
<td>&nbsp;</td>
<tr>
<td>Bioscreen 100-well Honeycomb plate</td>
<td>Tool</td>
<td>Growth Curves USA</td>
<td>9502550</td>
<td>&nbsp;</td>		</tbody>
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quantifying yeast chronological life span by outgrowth of aged cells

芽殖酵母<em&gt;酿酒酵母</em&gt;已被证明是一个重要的模式生物，在衰老研究领域<sup&gt; 1</sup&gt;。复制和时间顺序寿命是用来研究在酵母老化的两个既定的范式。复制性衰老定义为一个单一的酵母母细胞在衰老之前产生的子细胞的数量，按时间顺序老化时间细胞的长度定义能够生存在一个不分裂的，静止的状态<sup&gt; 2</sup&gt;。我们已经开发出一种高通量的方法按时间顺序寿命的定量测量。此方法涉及定义介质在搅拌下，在恒定温度和老化细胞。每个年龄点，一个细胞亚群是从老龄化的文化，并接种到丰富的生长介质。一个高分辨率的生长曲线，然后获得这种使用BIOSCREEN彗星的MBR机的老化细胞亚群。一个算法，然后应用，确定可行的细胞在每个分人口的相对比例，在每个年龄点的生长动力学。这种方法需要更少的时间比其他时间顺序寿命检测和资源，同时保持重现性和精密度。这个实验的高通量的性质，应允许进行大规模的遗传和化学的屏幕，在更复杂的生物进一步检测，以确定新型长寿修饰符。

Watch this Scientific Journal Video about Quantifying Yeast Chronological Life Span by Outgrowth of Aged Cells at JoVE.com

Quantifying Yeast Chronological Life Span by Outgrowth of Aged Cells

在酵母老化的年代，是指与固定相相关的细胞活力的损失。在这里，我们描述了一个高通量的方法，定量确定酵母按​​时间顺序寿命。

量化老化细胞生长的酵母年谱跨度

The budding yeast Saccharomyces cerevisiae has proven to be an important model organism in the field of aging research 1. The replicative and ...

Research

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Aging is a degenerative process characterized by a progressive deterioration of cellular components and organelles resulting in mortality. The nematode ...

科研

生物学

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在芽殖酵母中的自我复制寿命测量

Aging is a degenerative process characterized by a progressive deterioration of cellular components and organelles resulting in mortality. The budding ...

Purification of Mitochondria from Yeast Cells

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Mitochondria are the main site of ATP production during aerobic metabolism in eukaryotic non-photosynthetic cells1. These complex organelles also play ...

Measuring Caenorhabditis elegans Life Span in 96 Well Microtiter Plates

Measuring Caenorhabditis elegans Life Span in 96 Well Microtiter Plates

测量线虫寿命在96孔板

Lifespan is a biological process regulated by several genetic pathways. One strategy to investigate the biology of aging is to study animals that harbor ...

Quantitative Live Cell Fluorescence-microscopy Analysis of Fission Yeast

裂殖酵母的活细胞定量荧光显微镜分析

Several microscopy techniques are available today that can detect a specific protein within the cell. During the last decade live cell imaging using ...

Temporal Quantification of MAPK Induced Expression in Single Yeast Cells

在单酵母细胞MAPK诱导表达的时空定量

The quantification of gene expression at the  single cell level uncovers novel regulatory mechanisms obscured in measurements  performed at the population ...

In Vivo 4-Dimensional Tracking of Hematopoietic Stem and Progenitor Cells in Adult Mouse Calvarial Bone Marrow

In Vivo 4-Dimensional Tracking of Hematopoietic Stem and Progenitor Cells in Adult Mouse Calvarial Bone Marrow

在造血干细胞和祖细胞的成年小鼠颅骨骨髓体内四维跟踪

Through a delicate balance between quiescence and proliferation, self renewal and production of differentiated progeny, hematopoietic stem cells (HSCs) ...

Generation and Culture of Blood Outgrowth Endothelial Cells from Human Peripheral Blood

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Historically, the limited availability of primary endothelial cells from patients with vascular disorders has hindered the study of the molecular ...

Tracking and Quantifying Developmental Processes in C. elegans Using Open-source Tools

Tracking and Quantifying Developmental Processes in C. elegans Using Open-source Tools

跟踪和量化的发育过程<em&gt;℃。线虫</em&gt;使用开源工具

Quantitatively capturing developmental processes is crucial to derive mechanistic models and key to identify and describe mutant phenotypes. Here ...

Detection of Modified Forms of Cytosine Using Sensitive Immunohistochemistry

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