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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 |
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论文标题：Rapid and gentle hydrogel encapsulation of living organisms enables long-term microscopy over multiple hours
期刊：
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：
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，并能兼容使用明升手机处理手段，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.
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期刊介绍： () 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.
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