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秀丽隐杆线虫感觉神经元的染料填充条件。

Conditions for dye-filling of sensory neurons in Caenorhabditis elegans.

机构信息

Department of Biosciences and Nutrition, Karolinska Institutet, Huddinge, Sweden.

出版信息

J Neurosci Methods. 2010 Apr 30;188(1):58-61. doi: 10.1016/j.jneumeth.2010.02.003. Epub 2010 Feb 10.

Abstract

Dye-filling is a common method used to stain Caenorhabditis elegans sensory neurons in vivo. While the amphids and phasmids are easy to stain, a subset of sensory neurons, the IL2 neurons, are difficult to stain reproducibly. Here we examined the conditions under which the IL2 neurons take up the lipophilic fluorescent dye DiI. We find that IL2 dye-filling depends on salt concentration, but not osmolarity. Low salt prior and during incubation is important for efficient dye uptake. Additional parameters that affect dye-filling are the speed of shaking during incubation and the addition of detergents. Our modified dye-filling procedure provides a reliable method to distinguish mutant alleles that stain amphids and phasmids, IL2 neurons, or both. An additional benefit is that it can also stain the excretory duct. The method allows genetic screens to be performed to identify mutants that selectively affect only one of the sensory structures or the excretory duct.

摘要

染色是一种常用的方法,用于活体染色秀丽隐杆线虫的感觉神经元。尽管触角和尾感器很容易染色,但感觉神经元的一个亚群,IL2 神经元,很难重复染色。在这里,我们检查了 IL2 神经元摄取亲脂性荧光染料 DiI 的条件。我们发现 IL2 染色取决于盐浓度,但与渗透压无关。孵育前和孵育期间低盐是有效摄取染料的关键。影响染色的其他参数是孵育过程中摇晃的速度和去污剂的添加。我们改进的染色程序提供了一种可靠的方法来区分突变等位基因,这些等位基因可以染色触角和尾感器、IL2 神经元或两者。另一个好处是它还可以染色排泄管。该方法可用于进行遗传筛选,以鉴定仅选择性影响一种感觉结构或排泄管的突变体。

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