期刊
ANGEWANDTE CHEMIE-INTERNATIONAL EDITION
卷 56, 期 17, 页码 4724-4728出版社
WILEY-V C H VERLAG GMBH
DOI: 10.1002/anie.201612112
关键词
cell fixation; glutaraldehyde; microscopy; mitochondrial motion; photocleavage
资金
- leading-edge cluster Forum Organic Electronics as part of the High-Tech Strategy for Germany of the Federal Ministry of Education and Research [FKZ 13N11701]
- collaborative consortium MorphiQuant-3D [FKZ 13GW0044]
- Chemical Industry Fund of the German Chemical Industry Association (VCI)
Chemical fixation of living cells for microscopy is commonly achieved by crosslinking of intracellular proteins with dialdehydes prior to examination. We herein report a photocleavable protecting group for glutaraldehyde that results in a light-triggered and membrane-permeable fixative, which is nontoxic prior to photocleavage. Lipophilic ester groups allow for diffusion across the cell membrane and intracellular accumulation after enzymatic hydrolysis. Irradiation with UV light releases glutaraldehyde. The in situ generated fixative crosslinks intracellular proteins and preserves and stabilizes the cell so that it is ready for microscopy. In contrast to conventional glutaraldehyde fixation, tissue autofluorescence does not increase after fixation. Caged glutaraldehyde may in future enable functional experiments on living cells under a light microscope in which events of interest can be stopped in spatially confined volumes at defined time points. Samples with individually stopped events could then later be analyzed in ultrastructural studies.
作者
我是这篇论文的作者
点击您的名字以认领此论文并将其添加到您的个人资料中。
推荐
暂无数据