Name: yeast colony embedding method
Uploaded: 2011-03-22T11:00:00
Description: Watch this Scientific Journal Video about Yeast Colony Embedding Method at JoVE.com

研究是由美国国立卫生研究院1R15GM094770。

1。殖民地隔离和固定<ol><li>指定的时间孵育300琼脂培养基上菌落（一个孤立的殖民地应在直径1-2毫米）。 </li><li>删除殖民地（正面朝上）及相关的媒介，使用一个狭窄的铲。 </li><li>将数滴2％琼脂42 ° C的载玻片，使用琼脂面对1毫升pipetman提示，并立即将殖民地之前巩固。 </li><li>置于42℃菌落数滴2％琼脂，并允许以巩固。 </li><li>穿手套的所有步骤，通过本协议的其余部分</li><li>修剪块刀片，并?...

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	<li>Kimmel, A. R., Firtel, R. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Breaking+symmetries:+regulation+of+Dictyostelium+development+through+chemoattractant+and+morphogen+signal-response.">Breaking symmetries: regulation of Dictyostelium development through chemoattractant and morphogen signal-response.</a> Curr Opin Genet Dev. 14, 540-540 (2004).</li><li>Palkova, Z., Vachova, L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Life+within+a+community:+benefit+to+yeast+long-term+survival.">Life within a community: benefit to yeast long-term survival.</a> FEMS Microbiol Rev. 30, 806-806 (2006).</li><li>Shapiro, J., Vachova, L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+significances+of+bacterial+colony+patterns.">The significances of bacterial colony patterns.</a> Bioessays. 17, 597-597 (1995).</li><li>Shapiro, J., Vachova, L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Thinking+about+bacterial+populations+as+multicellular+organisms.">Thinking about bacterial populations as multicellular organisms.</a> Annu Rev Microbiol. 52, 81-81 (1998).</li><li>Engelberg, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Multicellular+stalk-like+structures+in+Saccharomyces+cerevisiae.">Multicellular stalk-like structures in Saccharomyces cerevisiae.</a> J Bacteriol. 180, 3992-3992 (1998).</li><li>Scherz, R., Shinder, V., Engelberg, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Anatomical+analysis+of+Saccharomyces+cerevisiae+stalk-like+structures+reveals+spatial+organization+and+cell+specialization.">Anatomical analysis of Saccharomyces cerevisiae stalk-like structures reveals spatial organization and cell specialization.</a> J Bacteriol. 183, 5402-5402 (2001).</li><li>Piccirillo, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+Rim101p/PacC+pathway+and+alkaline+pH+regulate+pattern+formation+in+yeast+colonies.">The Rim101p/PacC pathway and alkaline pH regulate pattern formation in yeast colonies.</a> Genetics. 184, 707-707 (2010).</li><li>Sniegowski, P. D., Dombrowski, P. G., Fingerman, E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Saccharomyces+cerevisiae+and+Saccharomyces+paradoxus+coexist+in+a+natural+woodland+site+in+North+America+and+display+different+levels+of+reproductive+isolation+from+European+conspecifics.">Saccharomyces cerevisiae and Saccharomyces paradoxus coexist in a natural woodland site in North America and display different levels of reproductive isolation from European conspecifics.</a> FEMS Yeast Res. 1, 299-299 (2002).</li><li>Piccirillo, S., Honigberg, S. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Sporulation+patterning+and+invasive+growth+in+wild+and+domesticated+yeast+colonies.">Sporulation patterning and invasive growth in wild and domesticated yeast colonies.</a> Res Microbiol. 161, 390-390 (2002).</li><li>Vachova, L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Architecture+of+developing+multicellular+yeast+colony:+spatio-temporal+expression+of+Ato1p+ammonium+exporter.">Architecture of developing multicellular yeast colony: spatio-temporal expression of Ato1p ammonium exporter.</a> Environ Microbiol. , (2009).</li></ol>

方法揭示了殖民地的内部结构。由于该方法是有效S 的范围内确定的细胞类型的模式酵母株不同的菌落形态，而且在一个相关的物种S. paradoxus 5，该方法也有可能工作的真菌和其他微生物的广泛。 方法的成功的一个关键的步骤是，以确保整个殖民地，包括殖民地的顶部，是在整个协议的琼脂包裹。无论是殖民地完全包裹在琼脂可以切片光镜和甲苯胺蓝染?...

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yeast colony embedding method

不同类型的细胞在胚胎的图案是一个后生发展的关键机制。殖民地和生物膜的微生物，如社区也显示细胞类型的模式。例如，在酵母S.的酵母 ，孢子细胞和假菌丝细胞不是均匀分布在菌落。图案和背后的分子机制，这些模式的功能的重要性仍然知之甚少。 调查真菌菌落的细胞类型模式的一个挑战是，不像后生动物的组织，细胞在殖民地比较弱连接到另一个。特别是，真菌菌落不包含在大多数组织中发现的细胞外基质的广泛的同一水平。在这里，我们报告上嵌入和切片酵母殖民地，揭示了在这些殖民地的细胞类型的内部模式的方法。该方法可用于准备厚切片，光镜和超薄切片（0.1μ）适合透射电子显微镜（0.5μ）有用。 ASCI和假菌丝细胞可以很容易地区别于卵形酵母细胞，通过光镜，而这些细胞的内部结构可以由EM观察。 该方法是基于对菌落周围的琼脂，渗透Spurr的培养基，然后切片。菌落直径在1-2毫米的范围内适用于本议定书。此外，以可视化的内部殖民地，该方法允许侵入底层琼脂的殖民地地区的可视化。

Watch this Scientific Journal Video about Yeast Colony Embedding Method at JoVE.com

Yeast Colony Embedding Method

嵌入酵母菌菌落，允许光学和电子显微镜切片的一种方法。此协议允许孢子细胞的分布和假菌丝细胞内提供了一个新的工具，对理解组织类型的细胞内一种真菌社会的殖民地的决心。

酵母殖民地嵌入方法

Patterning of  different cell types in embryos is a key mechanism in metazoan development. Communities of microorganisms, such as colonies and biofilms ...

Research

JoVE Journal

Biology

Plastic Embedding and Sectioning of Xenopus laevis Embryos

塑料嵌入和非洲爪蟾胚胎切片

科研

生物学

Measuring Replicative Life Span in the Budding Yeast

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

从酵母细胞线粒体的纯化

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

High-throughput Yeast Plasmid Overexpression Screen

高通量酵母质粒过表达屏幕

The budding yeast, Saccharomyces cerevisiae, is a powerful model system for defining fundamental mechanisms of many important cellular processes, ...

A Quantitative Assay for Insulin-expressing Colony-forming Progenitors

一个定量测定胰岛素表达细胞集落形成先觉

The field of pancreatic stem and progenitor cell biology has been hampered by a lack of in vitro functional and quantitative assays that allow for the ...

Cryosectioning Yeast Communities for Examining Fluorescence Patterns

检查荧光模式Cryosectioning酵母社区

Microbes typically live  in communities. The spatial organization  of cells within a community is believed to impact the survival and function of the  ...

The Soft Agar Colony Formation Assay

软琼脂集落形成分析

Anchorage-independent growth is the ability of transformed cells to grow independently of a solid surface, and is a hallmark of carcinogenesis. The soft ...

Surface Spreading and Immunostaining of Yeast Chromosomes

表面扩散和酵母染色体免疫染色

The small size of nuclei of the budding yeast Saccharomyces cerevisiae limits the utility of light microscopy for analysis of the subnuclear distribution ...

ODELAY: A Large-scale Method for Multi-parameter Quantification of Yeast Growth

ODELAY：用于酵母生长多参数定量的大规模方法

Growth phenotypes of microorganisms are a strong indicator of their underlying genetic fitness and can be segregated into 3 growth regimes: lag-phase, ...

Pancreatic Islet Embedding for Paraffin Sections

胰岛嵌段石蜡切片

Experiments using isolated pancreatic islets are important for diabetes research, but islets are expensive and of limited abundance. Islets contain a ...