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Cell fixation by light-triggered release of glutaraldehyde

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 non...

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Main Authors: Schelkle, Korwin (Author) , Schmidt, Christopher (Author) , Yserentant, Klaus (Author) , Bender, Markus (Author) , Wacker, Irene (Author) , Petzoldt, Martin (Author) , Hamburger, Manuel (Author) , Herten, Dirk-Peter (Author) , Wombacher, Richard (Author) , Schröder, Rasmus R. (Author) , Bunz, Uwe H. F. (Author)
Format: Article (Journal)
Language:English
Published: March 22, 2017
In: Angewandte Chemie. International edition
Year: 2017, Volume: 56, Issue: 17, Pages: 4724-4728
ISSN:1521-3773
DOI:10.1002/anie.201612112
Online Access:Verlag, Volltext: http://dx.doi.org/10.1002/anie.201612112
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Author Notes:Korwin M. Schelkle, Christopher Schmid, Klaus Yserentant, Markus Bender, Irene Wacker, Martin Petzoldt, Manuel Hamburger, Dirk-Peter Herten, Richard Wombacher, Rasmus R. Schröder, and Uwe H.F. Bunz
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Summary: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.
Item Description:Gesehen am 18.04.2018
Physical Description:Online Resource
ISSN:1521-3773
DOI:10.1002/anie.201612112

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