CLASI-FISH: principles of combinatorial labeling and spectral imaging.

CLASI-FISH: principles of combinatorial labeling and spectral imaging.
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DOI:
10.1016/j.syapm.2012.03.004
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发表时间:
2012-12
影响因子:
3.4
通讯作者:
Borisy GG
Borisy GG
中科院分区:
生物学2区
文献类型:
--
作者:
Valm AM;Mark Welch JL;Borisy GG

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尽管微生物群落中存在的系统类型的数量可能有数百个或更多,但直到最近,荧光原位杂交还被用来标记单个样本中的最多少数几个不同的系统类型。我们最近开发了一种技术,Clasi-FISH用于组合标记和光谱成像-荧光原位杂交,以极大地扩大单个鱼类实验中可区分的分类群的数量。Clasi技术涉及用探针和光谱成像的组合来标记感兴趣的微生物,以允许使用具有高度重叠的激发和发射光谱的荧光团。在这里,我们介绍了类鱼的基本原理和理论,以及进行类鱼实验的一些指导方针。我们还包括创建荧光光谱参考标准的协议,这是成功的Clasi-FISH的重要组成部分。
Although the number of phylotypes present in a microbial community may number in the hundreds or more, until recently, fluorescence in situ hybridization has been used to label, at most, only a handful of different phylotypes in a single sample. We recently developed a technique, CLASI-FISH for Combinatorial Labeling and Spectral Imaging – Fluorescence in situ Hybridization, to greatly expand the number of distinguishable taxa in a single FISH experiment. The CLASI technique involves labeling microbes of interest with combinations of probes coupled with spectral imaging to allow the use of fluorophores with highly overlapping excitation and emission spectra. Here, we present the basic principles and theory of CLASI-FISH along with some guidelines for performing CLASI-FISH experiments. We further include a protocol for creating fluorescence spectral reference standards, a vital component of successful CLASI-FISH.
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