Cell Fixation by Light‐Triggered Release of Glutaraldehyde. Issue 17 (22nd March 2017)

• Record Type: Journal Article
• Authors: Schelkle, Korwin M.; Schmid, Christopher; Yserentant, Klaus; Bender, Markus; Wacker, Irene; Petzoldt, Martin; Hamburger, Manuel; Herten, Dirk‐Peter; Wombacher, Richard; Schröder, Rasmus R.; Bunz, Uwe H. F.
• Is Part Of: Angewandte Chemie international edition. Volume 56:Issue 17(2017)
• Page Start: 4724
• Page End: 4728
• Publication Date: 2017-03-22
• Subjects: cell fixation -- glutaraldehyde -- microscopy -- mitochondrial motion -- photocleavage; Chemistry -- Periodicals; 540
• DOI: 10.1002/anie.201612112
• Languages: English
• ISSNs: 1433-7851

Abstract

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. Abstract : Cellfi‐Flash : A new fixative for light microscopy of living cells was obtained by functionalizing glutaraldehyde with a photocleavable protecting group. Ester substituents ensure that the masked compound can enter the cell and accumulate therein after esterase‐mediated hydrolysis. Incubated cells are instantly fixed and ready for microscopy upon irradiation, whereas conventional aldehyde fixation is diffusion‐controlled.