作者:Kyra Burnett, Eric Edsinger, Dirk R. Albrecht 来源:Communications Biology 发布时间:2018/10/10 9:56:35
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用水凝胶快速且温和地包封生物体来进行长时间的显微镜观察

论文标题:Rapid and gentle hydrogel encapsulation of living organisms enables long-term microscopy over multiple hours

期刊:Communications Biology

作者:Kyra Burnett, Eric Edsinger, Dirk R. Albrecht

发表时间:2018/06/21

数字识别码:10.1038/s42003-018-0079-6

原文链接:

https://www.nature.com/articles/s42003-018-0079-6?utm_source=Other_website&utm_medium=Website_links&utm_content=RenLi-MixedBrand-multijournal-Multidisciplinary-China&utm_campaign=ORG_USG_JRCN_RL_article_promotion_sciencenet_Oct_2nd

对生物体做高分辨度成像时需要对生物体做有效且无害的固定。如果实验时间较长,就更需要降低对生物体的干扰。在《通讯-生物学》发表的一篇名为Rapid and gentle hydrogel encapsulation of living organisms enables long-term microscopy over multiple hours的文章里,来自美国伍斯特理工学院的Dirk R. Albrecht的团队尝试用一个简单和经济的方法,通过光交联聚乙二醇(PEG)水凝胶来封装处于任意发育阶段的小型动物,并成功地对它们作温和的固定。被固定的动物在这种水分充足的环境里能够保持其原来形貌,并能兼容使用化学处理手段,光刺激手段,和使用光片显微镜。

通过此方法,作者对秀丽隐杆线虫进行了长时间了3D神经反应成像,并在24小时后成功恢复活性有机体,还对较大的鱿鱼幼体进行了成像。

作者研究了一系列的水凝胶和光照条件,来检测固定效果,最终确定了适合于高分辨率单细胞成像且不会造成麻痹的条件。总而言之,PEG水凝胶包封法提供了快速,通用且温和的方式来固定小型生物活体,从酵母菌到斑马鱼均可承受数小时的连续观察。

图1:PEG水凝胶能够快速简单地把生物固定。

摘要:Imaging living organisms at high spatial resolution requires effective and innocuous immobilization. Long-term imaging places further demands on sample mounting with minimal perturbation of the organism. Here we present a simple, inexpensive method for rapid encapsulation of small animals of any developmental stage within a photo-crosslinked polyethylene glycol (PEG) hydrogel, gently restricting movement within their confined spaces. Immobilized animals maintain their original morphology in a hydrated environment compatible with chemical treatment, optical stimulation, and light-sheet microscopy. We demonstrate prolonged three-dimensional imaging of neural responses in the nematode Caenorhabditis elegans, recovery of viable organisms after 24 h, and imaging of larger squid hatchlings. We characterize a range of hydrogel and illumination conditions for immobilization quality, and identify paralytic-free conditions suitable for high-resolution single-cell imaging. Overall, PEG hydrogel encapsulation provides fast, versatile, and gentle mounting of small living organisms, from yeast to zebrafish, for continuous observation over hours.

阅读论文全文请访问:https://www.nature.com/articles/s42003-018-0079-6?utm_source=Other_website&utm_medium=Website_links&utm_content=RenLi-MixedBrand-multijournal-Multidisciplinary-China&utm_campaign=ORG_USG_JRCN_RL_article_promotion_sciencenet_Oct_2nd

期刊介绍:Communications Biology (https://www.nature.com/commsbio/) is an open access journal from Nature Research publishing high-quality research, reviews and commentary in all areas of the biological sciences. Research papers published by the journal represent significant advances bringing new biological insight to a specialized area of research.

(来源:科学网)

 
 
 
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