A low percent ethanol method for immobilizing planarians.
A low percent ethanol method for immobilizing planarians.
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低百分比乙醇方法固定平面的人。
DOI：
10.1371/journal.pone.0015310
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发表时间：
2010-12-14
期刊：
影响因子：
3.7
通讯作者：
Beane WS
中科院分区：
文献类型：
--
作者：
Stevenson CG;Beane WS
近年来，涡虫已成为研究成体干细胞、再生和极性的热门模型系统。该系统对本科生和研究生的研究实验室都很有吸引力，因为涡虫菌落成本低，易于维护。此外，原位杂交、免疫荧光和rna干扰（RNAi）基因敲除技术也已发展用于涡虫研究。然而，活体蠕虫的成像（特别是在高倍放大下）是困难的，因为动物有强烈的恐光性；它们很快地从光源移开并移出画面。目前可用的抑制涡虫运动的方法包括RNAi注射和暴露在低温下。前者是劳动和时间密集，而后者排除了许多荧光报告染料的使用。在这里，我们报告了一种简单、廉价和可逆的方法来固定涡虫进行实时成像。我们的数据表明，用3%乙醇（EtOH）处理1小时就足以抑制地中海施米德（Schmidtea mediterranea）典型体型（4-6毫米）的精细和粗大运动，并在3-4小时内完全恢复运动。重要的是，即使在反复暴露后，EtOH处理也不会干扰再生，也不会溶解上皮细胞（通过H&E染色检测）。我们证明，短时间暴露于低浓度的EtOH是一种快速有效的固定涡虫的方法，这种方法很容易适用于各种大小的涡虫，并将增加涡虫研究人员实时成像分析的可及性。
Planarians have recently become a popular model system for the study of adult stem cells, regeneration and polarity. The system is attractive for both undergraduate and graduate research labs, since planarian colonies are low cost and easy to maintain. Also in situ hybridization, immunofluorescence and RNA-interference (RNAi) gene knockdown techniques have been developed for planarian studies. However, imaging of live worms (particularly at high magnifications) is difficult because animals are strongly photophobic; they quickly move away from light sources and out of frame. The current methods available to inhibit movement in planarians include RNAi injection and exposure to cold temperatures. The former is labor and time intensive, while the latter precludes the use of many fluorescent reporter dyes. Here, we report a simple, inexpensive and reversible method to immobilize planarians for live imaging. Our data show that a short 1 hour treatment with 3% ethanol (EtOH) is sufficient to inhibit both the fine and gross movements of Schmidtea mediterranea planarians, of the typical size used (4–6 mm), with full recovery of movement within 3–4 hours. Importantly, EtOH treatment did not interfere with regeneration, even after repeated exposure, nor lyse epithelial cells (as assayed by H&E staining). We demonstrate that a short exposure to a low concentration of EtOH is a quick and effective method of immobilizing planarians, one that is easily adaptable to planarians of all sizes and will increase the accessibility of live imaging assays to planarian researchers.
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影响因子：
3.8
作者：
Nogi T;Zhang D;Chan JD;Marchant JS
通讯作者：
Marchant JS
影响因子：
3.6
作者：
Chakrabarti, A;Schatten, H;Taylor, M
通讯作者：
Taylor, M
DOI：
10.1002/neu.20290
发表时间：
2006-08-01
期刊：
JOURNAL OF NEUROBIOLOGY
影响因子：
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作者：
Hicks, Caitlin;Sorocco, Debra;Levin, Michael
通讯作者：
Levin, Michael
影响因子：
2.9
作者：
Cebria, Francesc
通讯作者：
Cebria, Francesc
DOI：
10.1016/s0305-0491(97)00328-3
发表时间：
1998-02-01
影响因子：
2.2
作者：
McGee, C;Wisdom, GB;Walker, B
通讯作者：
Walker, B