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Cell Fixation by Light‐Triggered Release of Glutaraldehyde
Author(s) -
Schelkle Korwin M.,
Schmid Christopher,
Yserentant Klaus,
Bender Markus,
Wacker Irene,
Petzoldt Martin,
Hamburger Manuel,
Herten DirkPeter,
Wombacher Richard,
Schröder Rasmus R.,
Bunz Uwe H. F.
Publication year - 2017
Publication title -
angewandte chemie
Language(s) - English
Resource type - Journals
eISSN - 1521-3757
pISSN - 0044-8249
DOI - 10.1002/ange.201612112
Subject(s) - glutaraldehyde , fixative , intracellular , biophysics , fixation (population genetics) , chemistry , membrane , cell membrane , autofluorescence , microscopy , cell , biochemistry , chromatography , biology , cytoplasm , fluorescence , pathology , medicine , physics , quantum mechanics , gene
Abstract 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.