Rapid and gentle hydrogel encapsulation of living organisms enables long-term microscopy over multiple hours.
Rapid and gentle hydrogel encapsulation of living organisms enables long-term microscopy over multiple hours.
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DOI：
10.1038/s42003-018-0079-6
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发表时间：
2018
影响因子：
5.9
通讯作者：
Albrecht DR
中科院分区：
文献类型：
--
作者：
Burnett K;Edsinger E;Albrecht DR
以高空间分辨率成像生物体需要有效和无害的固定。长期成像进一步要求在对生物体的干扰最小的情况下安装样品。在这里，我们提出了一种简单，廉价的方法，用于快速封装的任何发育阶段的小动物在光交联聚乙二醇（PEG）水凝胶，轻轻地限制其有限的空间内的运动。固定动物保持其原始形态在水合环境中与化学处理，光学刺激，和光片显微镜兼容。我们展示了线虫的神经反应的长时间三维成像，24小时后恢复的活生物体，和较大的鱿鱼孵化的成像。我们表征了一系列的水凝胶和照明条件的固定质量，并确定适合高分辨率单细胞成像的无麻痹条件。总的来说，PEG水凝胶封装提供了快速，通用，温和的安装小的活生物体，从酵母到斑马鱼，连续观察数小时。Kyra Burnett等人提出了一种简单而经济的方法，将小型活生物体封装在光交联聚乙二醇水凝胶中，用于长期显微镜检查。这种方法提供了一个快速和温和的安装连续成像数小时，并与光片显微镜和光遗传学刺激。
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. Kyra Burnett et al. present a simple and economical method to encapsulate small living organisms for long-term microscopy in a photo-crosslinked polyethylene glycol hydrogel. This method provides a fast and gentle mounting for continuous imaging over hours, and works with light-sheet microscopy and optogenetic stimulation.
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带有微流体学的秀丽隐杆线虫幼虫的长期高分辨率成像。
影响因子：
11.8
作者：
Keil W;Kutscher LM;Shaham S;Siggia ED
通讯作者：
Siggia ED
影响因子：
--
作者：
Fang-Yen, Christopher;Gabel, Christopher V.;Samuel, Aravinthan D. T.;Bargmann, Cornelia I.;Avery, Leon
通讯作者：
Avery, Leon
影响因子：
--
作者：
Goodman, Miriam B.;Lindsay, Theodore H.;Lockery, Shawn R.;Richmond, Janet E.
通讯作者：
Richmond, Janet E.
影响因子：
9
作者：
Hockaday LA;Kang KH;Colangelo NW;Cheung PY;Duan B;Malone E;Wu J;Girardi LN;Bonassar LJ;Lipson H;Chu CC;Butcher JT
通讯作者：
Butcher JT
影响因子：
6.1
作者：
Albrecht, DR;Tsang, VL;Bhatia, SN
通讯作者：
Bhatia, SN